Compounds and compositions as channel activating protease inhibitors

ABSTRACT

The invention provides compounds and pharmaceutical compositions thereof, which are useful for modulating channel activating proteases, and methods for, using such compounds to treat, ameliorate or prevent a condition associated with a channel activating protease, including but not limited to prostasin, PRSS22, TMPRSS11 (e.g., TMPRSS11B, TMPRSS11E), TMPRSS2, TMPRSS3, TMPRSS4 (MTSP-2), matriptase (MTSP-1), CAP2, CAP3, trypsin, cathepsin A, or neutrophil elastase.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. provisional applicationsSer. No. 60/802,983, filed 23 May 2006; and Ser. No. 60/860,604, filed22 Nov. 2006. Each of these applications is incorporated herein byreference in its entirety.

TECHNICAL FIELD

The invention generally relates to channel activating protease (CAP)inhibitors.

BACKGROUND ART

Prostasin is a trypsin-like serine protease that is present in a varietyof mammalian tissues. It is a membrane anchored protease that isexpressed on the extra-cellular membrane of cells, but may also besecreted into body fluids such as semen, urine and airway surfaceliquid. Prostasin (PRSS8), together with proteases such as matriptase,CAP2, CAP3, trypsin, PRSS22, TMPRSS11, cathepsin A, and neutrophilelastase, may stimulate the activity of the amiloride-sensitiveepithelial sodium channel (ENaC). Inhibiting these enzymes may inducechanges in epithelial ion transport and therefore fluid homeostasisacross epithelial membranes. For example, CAP inhibition in the kidneyis thought to promote diuresis, whilst CAP inhibition in the airwayspromotes the clearance of mucus and sputum in lung. CAP inhibition inthe kidney may therefore be used therapeutically to treat hypertension.CAP inhibition in the airways prevents the stagnation of respiratorysecretions that otherwise tends to make sufferers vulnerable tosecondary bacterial infections.

DISCLOSURE OF THE INVENTION

The invention provides compounds, pharmaceutical compositions andmethods of using such compounds for modulating channel activatingproteases (CAP). For example, the compounds and compositions of theinvention may be used for modulating prostasin, PRSS22, TMPRSS11 (e.g.,TMPRSS11B, TMPRSS11E), TMPRSS2, TMPRSS3, TMPRSS4 (MTSP-2), matriptase(MTSP-1), CAP2, CAP3, trypsin, cathepsin A, and neutrophil elastase.

In one aspect, the present invention provides compounds of Formula (1):

and pharmaceutically acceptable salts, hydrates, solvates andstereoisomers thereof, wherein:

J is a 5-12 membered monocyclic or fused carbocyclic ring, aryl,heteroaryl or heterocyclic ring containing N, O and/or S;

R¹ is —(CR₂)_(l)—NR₂, —(CR₂)_(l)—NRC(═NR)—NR₂, —(CR₂)_(l)—C(═NR)—NR₂ ora 5-7 membered nitrogen-containing non-aromatic heterocyclic ring;

W—R² is a substituent at any position on ring A;

W is —O(CR₂)_(k)—, —S(CR₂)_(k)—, —S(O)(CR₂)_(k)—, —SO₂(CR₂)_(k)— or—OC(O)(CR₂)_(k)—;

R² is C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, R⁸, —CR¹⁰═CR¹⁰—R⁸, or

wherein ring E is an optionally substituted 5-7 membered monocyclic orfused carbocyclic or heterocyclic ring; or W—R² together form C₁₋₆alkyl, a 5-7 membered aryl or —OC(O)NR⁶R⁷;

Y is SO₂R³, —(CO)—NR—R³, —(CO)—O—R³, SO₂NR⁶R⁷ or C₁₋₆ alkyl;

R³ is C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, —(CR₂)_(l)—C₃₋₇ cycloalkylor —(CR₂)_(l)—R⁸;

R⁴ is H, C₁₋₆ alkyl, C₂₋₆ alkenyl, —CR¹⁰═CR¹⁰—R⁸, —CR[(CR₂)_(l)—R⁸]₂,C₂₋₆ alkynyl, —O—(CR₂)_(l)—R⁹, NR⁶R⁷,

or an optionally substituted 5-7 membered carbocyclic ring, heterocyclicring, aryl or heteroaryl; or R⁴ together with Y form an optionallysubstituted 5-12 membered non-aromatic heterocyclic ring;

R⁵ is halo, C₁₋₆ alkyl, C₁₋₆ alkoxy, OR⁹ or R⁹;

R⁶ and R⁷ are independently H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl or—(CR₂)_(l)—R⁸; or R⁶ and R⁷ together with N may form an optionallysubstituted 5-7 membered monocyclic or fused heterocyclic ring;

X, R⁸ and R⁹ are independently an optionally substituted 5-7 memberedcarbocyclic ring, heterocyclic ring, aryl or heteroaryl; or R⁹ may be Hor C₁₋₆ alkyl;

R¹⁰ is H or C₁₋₆ alkyl;

each R is H, C₁₋₆ alkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl, wherein a carbonmay optionally be substituted or replaced with NR, O or S;

i is 0-1;

k, l and m are independently 0-6;

n is 1-6; and

p is 0-3.

In compounds of Formula (1), R² may be an optionally substituted phenyl,thienyl, C₅₋₇ cycloalkyl (e.g., cyclohexyl or cyclopentyl), furanyl,piperidinyl, methylenecyclohexyl,

For example, R² substituents may be optionally substituted with halo,CF₃, C₁₋₆ alkyl, C₂₋₆ alkenyl, O—(CH₂)₀₋₄—R⁹.

In one embodiment, the invention provides compounds of Formula (2A) or(2B):

wherein Z is O or S;

R¹ is NH₂, —NHC(═NH)—NH₂ or —C(═NH)—NH₂;

W is —O(CH₂)_(k)— or —S(O)(CH₂)_(k)—;

R² is an optionally substituted phenyl, or W—R² together form C₁₋₆ alkylor an optionally substituted phenyl;

Y is SO₂R³ or —(CO)—O—R³;

R³ is C₁₋₆ alkyl, —(CH₂)_(l)-cyclopropyl or —(CH₂)_(l)—R⁸ wherein R⁸ isan optionally substituted phenyl;

R⁴ is an optionally substituted, phenyl, piperidinyl, C₅₋₇ cycloalkyl,cyclohexanol, imidazolyl, thienyl,

i and p are 0;

k is 1;

l is 0-1; and

m and n are independently 1-4.

In some examples, the compounds are of Formula (2A) or (2B) and R⁴ ispiperidinyl. In other examples, Z is O.

In compounds of Formula (2A) or (2B) where R² is phenyl or W—R² togetherform phenyl, each phenyl may optionally be substituted with halo.

In compounds of Formula (1), (2A) and (2B), W may be —O(CR₂)_(k)—,—S(CR₂)_(k)—, —S(O)(CR₂)_(k)—, —SO₂(CR₂)_(k)— or —OC(O)(CR₂)_(k)—; and kis 1.

In another embodiment, the invention provides compounds of Formula (3A)or (3B):

wherein R⁶ and R⁷ are independently H, C₁₋₆ alkyl or —(CR₂)_(l)—R⁸; orR⁶ and R⁷ together with N form an optionally substituted pyrrolidinyl,piperidinyl, morpholino, piperazinyl or diazepanyl;

R⁸ is an optionally substituted phenyl, furanyl, tetrahydrofuranyl,piperidinyl or thienyl; and

i and p are 0.

In some compounds of Formula (3A) or (3B), each optionally substitutedsubstituent may be substituted with NR⁹ ₂, halo, C₁₋₆ alkyl,—(CR₂)_(l)—COR⁹, —(CR₂)_(l)—R⁹, —(CR₂)_(l)—SO₂R⁹ or NRSO₂R⁹, wherein R⁹is H, C₁₋₆ alkyl, or an optionally substituted carbocyclic ring,heterocyclic ring, aryl or heteroaryl. For example, R⁹ may be phenyl,furanyl, tetrahydrofuranyl or thienyl, each of which may be optionallysubstituted by substituents described above.

In the compounds of the invention, R¹ may be —(CH₂)_(l)—NH₂,—(CH₂)_(l)—NHC(═NH)—NH₂ or —(CH₂)_(l)—C(═NH)—NH₂NH₂, wherein each l is0-1; or R¹ is piperidinyl. In some examples, each R group in (CR₂) is H.

In the compounds of the invention, Y may be SO₂R³, —(CO)—NH—R³ or—(CO)—O—R³; R³ is C₁₋₆ alkyl, —(CR₂)_(l)—C₃₋₇ cycloalkyl (e.g.,cyclopentyl, cyclohexyl and particularly cyclopropyl, each of which maybe optionally substituted), or —(CR₂)_(l)—R⁸ where R⁸ is an optionallysubstituted phenyl. Optional substituents for R³ and R⁸ include but arenot limited to halo, CF₃, C₁₋₆ alkyl, C₂₋₆ alkenyl or O—(CH₂)₀₋₄—R⁹.

In the compounds of the invention, R⁴ may be H, C₁₋₆ alkyl, C₂₋₆alkenyl, —NH₂, or an optionally substituted phenyl, phenoxy,piperidinyl, cyclohexyl, cyclohexanol, imidazolyl, thienyl,

or R⁴ together with Y form an optionally substituted pyrrolidinyl,pyrrolidinonyl, tetrahydroisoquinolinyl or tetrahydronapthalenyl. Eachoptionally substituted substituent may be substituted with halo, CF₃,OCF₃, C₁₋₆ alkyl, C₂₋₆ alkenyl, R⁸, O—(CH₂)₀₋₄—R⁹ or CO₂R⁹, wherein R⁹may be H, C1-6 alkyl, or phenyl.

In the compounds of the invention, -J-(R⁵)_(p) together may be

wherein Z is O or S;

Z¹, Z², Z³ or Z⁴ are independently N, CH, or C when attached to R⁵;

Z⁵, Z⁶ or Z⁷ are independently N, O, S, CH, or C when attached to R⁵;

p is 0-1; and

R⁵ is halo or C₁₋₆ alkyl.

In the compounds of the invention, J may be benzothiazolyl,benzoxazolyl, thiazolyl, or oxadiazolyl. In particular examples, J isbenzothiazolyl or benzoxazolyl.

In the compounds of the invention where R⁸ is a substituent, R⁸ may bean optionally substituted phenyl, C₅₋₇ cycloalkyl (e.g., cyclopentyl orcyclohexyl), piperidinyl, cyclohexanol, imidazolyl, thienyl, furanyl,

In the compounds of the invention, X may be cyclohexyl, phenyl orpiperidinyl, each of which is optionally substituted with C₁₋₆ alkyl,C₁₋₆ alkoxy, halo, or a combination thereof.

In another aspect, the present invention provides pharmaceuticalcompositions comprising a compound having Formula (1), (2A), (2B), (3A)or (3B), and a pharmaceutically acceptable excipient.

The invention also provides methods for modulating a channel activatingprotease, comprising administering to a system or a subject in needthereof, a therapeutically effective amount of a compound having Formula(1), (2A), (2B), (3A) or (3B), or pharmaceutically acceptable salts orpharmaceutical compositions thereof, thereby modulating said channelactivating protease.

In yet another aspect, the invention provides methods for ameliorating acondition mediated by a channel activating protease, comprisingadministering to a system or subject in need of such treatment aneffective amount of a compound having Formula (1), (2A), (2B), (3A) or(3B), or pharmaceutically acceptable salts or pharmaceuticalcompositions thereof, and optionally in combination with a secondtherapeutic agent, thereby treating said condition. Examples of a secondtherapeutic agent which may be used with the compounds of the inventioninclude but are not limited to an anti-inflammatory, bronchodilatory,antihistamine, anti-tussive, antibiotic or DNase.

Examples of channel activating protease which may be modulated using thecompounds of the invention include but are not limited to prostasin,PRSS22, TMPRSS11 (e.g., TMPRSS11B, TMPRSS11E), TMPRSS2, TMPRSS3, TMPRSS4(MTSP-2), matriptase (MTSP-1), CAP2, CAP3, trypsin, cathepsin A, orneutrophil elastase. In particular examples, the invention providesmethods for modulating prostasin, or methods for treating a conditionmediated by prostasin.

In the above methods for using the compounds of the invention, acompound having Formula (1), (2A), (2B), (3A) or (3B) may beadministered to a system comprising cells or tissues. For example, acompound having Formula (1), (2A), (2B), (3A) or (3B) may be contactedwith bronchial epithelial cells, which may be human cells. In otherembodiments, a compound having Formula (1), (2A), (2B), (3A) or (3B) maybe administered to a human or animal subject.

In one embodiment, the invention provides methods for ameliorating acondition associated with the movement of fluid across ion transportingepithelia or the accumulation of mucus and sputum in respiratorytissues, or a combination thereof. For example, the condition may becystic fibrosis, primary ciliary dyskinesia, lung carcinoma, chronicbronchitis, chronic obstructive pulmonary disease, asthma or arespiratory tract infection.

Furthermore, the present invention provides the use of a compound ofFormula (1), (2A), (2B), (3A) or (3B), or pharmaceutically acceptablesalts or pharmaceutical compositions thereof, and optionally incombination with a second therapeutic agent, for modulating a channelactivating protease (e.g., for inhibiting prostasin). The presentinvention also provides the use of a compound having Formula (1), (2A),(2B), (3A) or (3B), or pharmaceutically acceptable salts orpharmaceutical compositions thereof, and optionally in combination witha second therapeutic agent, in the manufacture of a medicament fortreating a condition mediated by a channel activating protease (e.g., aprostasin-mediated condition).

DEFINITIONS

“Alkyl” refers to a moiety and as a structural element of other groups,for example halo-substituted-alkyl and alkoxy, and may bestraight-chained or branched. An optionally substituted alkyl, alkenylor alkenyl as used herein may be optionally halogenated (e.g., CF₃), ormay have one or more carbons that is substituted or replaced with aheteroatom, such as NR, O or S (e.g., —OCH₂CH₂O—, alkylthiols,thioalkoxy, alkylamines, etc).

“Aryl” refers to a monocyclic or fused bicyclic aromatic ring containingcarbon atoms. For example, aryl may be phenyl or naphthyl. “Arylene”means a divalent radical derived from an aryl group.

“Heteroaryl” as used herein is as defined for aryl above, where one ormore of the ring members is a heteroatom. Examples of heteroarylsinclude but are not limited to pyridyl, indolyl, indazolyl,quinoxalinyl, quinolinyl, benzofuranyl, benzopyranyl, benzothiopyranyl,benzo[1,3]dioxole, imidazolyl, benzo-imidazolyl, pyrimidinyl, furanyl,oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, thienyl, etc.

A “carbocyclic ring” as used herein refers to a saturated or partiallyunsaturated, monocyclic, fused bicyclic or bridged polycyclic ringcontaining carbon atoms, which may optionally be substituted, forexample, with ═O. Examples of carbocyclic rings include but are notlimited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cyclopropylene, cyclohexanone, etc.

A “heterocyclic ring” as used herein is as defined for a carbocyclicring above, wherein one or more ring carbons is a heteroatom. Forexample, a heterocyclic ring may contain N, O, S, —N═, —S—, —S(O),—S(O)₂—, or —NR— wherein R may be hydrogen, C₁₋₄alkyl or a protectinggroup. Examples of heterocyclic rings include but are not limited tomorpholino, pyrrolidinyl, pyrrolidinyl-2-one, piperazinyl, piperidinyl,piperidinylone, 1,4-dioxa-8-aza-spiro[4,5]dec-8-yl, etc.

The terms “co-administration” or “combined administration” or the likeas used herein are meant to encompass administration of the selectedtherapeutic agents to a single patient, and are intended to includetreatment regimens in which the agents are not necessarily administeredby the same route of administration or at the same time.

The term “pharmaceutical combination” as used herein refers to a productobtained from mixing or combining active ingredients, and includes bothfixed and non-fixed combinations of the active ingredients. The term“fixed combination” means that the active ingredients, e.g. a compoundof Formula (1), (2A), (2B), (3A) or (3B) and a co-agent, are bothadministered to a patient simultaneously in the form of a single entityor dosage. The term “non-fixed combination” means that the activeingredients, e.g. a compound of Formula (1), (2A), (2B), (3A) or (3B)and a co-agent, are both administered to a patient as separate entitieseither simultaneously, concurrently or sequentially with no specifictime limits, wherein such administration provides therapeuticallyeffective levels of the active ingredients in the body of the patient.The latter also applies to cocktail therapy, e.g. the administration ofthree or more active ingredients.

The term “therapeutically effective amount” means the amount of thesubject compound that will elicit a biological or medical response in acell, tissue, organ, system, animal or human that is being sought by theresearcher, veterinarian, medical doctor or other clinician.

The term “administration” and or “administering” of the subject compoundshould be understood to mean as providing a compound of the inventionand prodrugs thereof, to the individual in need of treatment.

As used herein, the terms “treat”, “treating” and “treatment” refer to amethod of alleviating or abating a disease and/or its attendantsymptoms.

The term “prostasin” may also be referred to as: humanchannel-activating protease (hCAP); channel-activating protease-1; andPRSS8, MERPOPS ID S01.159.

MODES OF CARRYING OUT THE INVENTION

The invention provides compounds, pharmaceutical compositions andmethods of using such compounds for modulating channel activatingproteases (CAP).

In one aspect, the present invention provides compounds of Formula (1):

and pharmaceutically acceptable salts, hydrates, solvates andstereoisomers thereof, wherein:

J is a 5-12 membered monocyclic or fused carbocyclic ring, aryl,heteroaryl or heterocyclic ring containing N, O and/or S;

R¹ is —(CR₂)_(l)—NR₂, —(CR₂)_(l)—NRC(═NR)—NR₂, —(CR₂)_(l)—C(═NR)—NR₂ ora 5-7 membered nitrogen-containing non-aromatic heterocyclic ring;

W—R² is a substituent at any position on ring A;

W is —O(CR₂)_(k)—, —S(CR₂)_(k)—, —S(O)(CR₂)_(k)—, —SO₂(CR₂)_(k)— or—OC(O)(CR₂)_(k)—;

R² is C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, R⁸, —CR¹⁰═CR¹⁰—R⁸, or

wherein ring E is an optionally substituted 5-7 membered monocyclic orfused carbocyclic or heterocyclic ring; or W—R² together form C₁₋₆alkyl, a 5-7 membered aryl or —OC(O)NR⁶R⁷;

Y is SO₂R³, —(CO)—NR—R³, —(CO)—O—R³, SO₂NR⁶R⁷ or C₁₋₆ alkyl;

R³ is C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, —(CR₂)_(l)—C₃₋₇ cycloalkylor —(CR₂)_(l)—R⁸;

R⁴ is H, C₁₋₆ alkyl, C₂₋₆ alkenyl, —CR¹⁰═CR¹⁰—R⁸, —CR[(CR₂)_(l)—R⁸]₂,C₂₋₆ alkynyl, —O—(CR₂)_(l)—R⁹, NR⁶R⁷,

or an optionally substituted 5-7 membered carbocyclic ring, heterocyclicring, aryl or heteroaryl; or R⁴ together with Y form an optionallysubstituted 5-12 membered non-aromatic heterocyclic ring;

R⁵ is halo, C₁₋₆ alkyl, C₁₋₆ alkoxy, OR⁹ or R⁹;

R⁶ and R⁷ are independently H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl or—(CR₂)_(l)—R⁸; or R⁶ and R⁷ together with N may form an optionallysubstituted 5-7 membered monocyclic or fused heterocyclic ring;

X, R⁸ and R⁹ are independently an optionally substituted 5-7 memberedcarbocyclic ring, heterocyclic ring, aryl or heteroaryl; or R⁹ may be Hor C₁₋₆ alkyl;

R¹⁰ is H or C₁₋₆ alkyl;

each R is H, C₁₋₆ alkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl, wherein a carbonmay optionally be substituted or replaced with NR, O or S;

i is 0-1;

k, l and m are independently 0-6;

n is 1-6; and

p is 0-3.

In one embodiment, the compounds of the invention have Formula (2A) or(2B):

wherein Z is O or S;

R¹ is NH₂, —NHC(═NH)—NH₂ or —C(═NH)—NH₂;

W is —O(CH₂)_(k)— or —S(O)(CH₂)_(k)—;

R² is an optionally substituted phenyl, or W—R² together form C₁₋₆ alkylor an optionally substituted phenyl;

Y is SO₂R³ or —(CO)—O—R³;

R³ is C₁₋₆ alkyl, —(CH₂)_(l)-cyclopropyl or —(CH₂)_(l)—R⁸ wherein R⁸ isan optionally substituted phenyl;

R⁴ is an optionally substituted, phenyl, piperidinyl, C₅₋₇ cycloalkyl,cyclohexanol, imidazolyl, thienyl,

i and p are 0;

k is 1;

l is 0-1; and

m and n are independently 1-4.

In another embodiment, the compounds of the invention have Formula (3A)or (3B):

wherein R⁶ and R⁷ are independently H, C₁₋₆ alkyl or —(CR₂)_(l)—R⁸; orR⁶ and R⁷ together with N form an optionally substituted pyrrolidinyl,piperidinyl, morpholino, piperazinyl or diazepanyl;

R⁸ is an optionally substituted phenyl, furanyl, tetrahydrofuranyl,piperidinyl or thienyl; and

i and p are 0.

In some embodiments, X, R⁸ and R⁹ may be an optionally substituted C₃₋₇cycloalkyl.

In each of the above formula, R¹ may be NR′R″, NH—C(NR′R″)═NH,NH—C(NHR′)═NR″, NH—C(R′)═NR″, S—C(NR′R″)—NH, S—C(NHR′)—NR″, C(NR′R″)═NH,C(NHR′)═NR″ or CR═NR″; where R′R″ are the same or different and are H,C₁₋₆ alkyl, C₁₋₃ arylalkyl, aryl or where R′R″ forms anitrogen-containing nonaromatic heterocyclic cyclic ring.

The compounds and compositions of the invention may be useful formodulating a channel activating protease. Examples of channel activatingproteases which may be modulated using the compounds and compositions ofthe invention include but are not limited to prostasin, PRSS22, TMPRSS11(e.g., TMPRSS11B, TMPRSS11E), TMPRSS2, TMPRSS3, TMPRSS4 (MTSP-2),matriptase (MTSP-1), CAP2, CAP3, trypsin, cathepsin A, or neutrophilelastase. The novel compounds of this invention may also inhibit theactivity of proteases that stimulate the activity of ion channels, suchas the epithelial sodium channel, and may be useful in the treatment ofCAP-associated diseases.

Pharmacology and Utility

Compounds of the invention modulate the activity of channel activatingprotease, for example, trypsin-like serine proteases such as prostasin,and as such, are useful for treating diseases or disorders in whichprostasin contributes to the pathology and/or symptomology of thedisease.

Diseases mediated by inhibition of a channel activating protease, forexample, by a trypsin-like serine protease such as prostasin, includediseases associated with the regulation of fluid volumes acrossepithelial membranes. For example, the volume of airway surface liquidis a key regulator of mucociliary clearance and the maintenance of lunghealth. The inhibition of a channel activating protease will promotefluid accumulation on the mucosal side of the airway epithelium, therebypromoting mucus clearance and preventing the accumulation of mucus andsputum in respiratory tissues (including lung airways). Such diseasesinclude respiratory diseases such as cystic fibrosis, primary ciliarydyskinesia, chronic bronchitis, chronic obstructive pulmonary disease(COPD), asthma, respiratory tract infections (acute and chronic; viraland bacterial) and lung carcinoma. Diseases mediated by inhibition ofchannel activating proteases also include diseases other thanrespiratory diseases that are associated with abnormal fluid regulationacross an epithelium, perhaps involving abnormal physiology of theprotective surface liquids on their surface, for example xerostomia (drymouth) or keratoconjunctivitis sire (dry eye). Furthermore, CAPregulation of ENaC in the kidney could be used to promote diuresis andthereby induce a hypotensive effect.

Chronic obstructive pulmonary disease includes chronic bronchitis ordyspnoea associated therewith, emphysema, as well as exacerbation ofairways hyper-reactivity consequent to other drug therapy, in particularother inhaled drug therapy. The invention is also applicable to thetreatment of bronchitis of whatever type or genesis including, forexample, acute, arachidic, catarrhal, croupus, chronic or phthinoidbronchitis.

The compounds of the invention may be used for the treatment of asthma,including but not limited to intrinsic (non-allergic) asthma andextrinsic (allergic) asthma, mild asthma, moderate asthma, severeasthma, bronchitic asthma, exercise-induced asthma, occupational asthmaand asthma induced following bacterial infection. Treatment of asthma isalso to be understood as embracing treatment of subjects of, forexample, less than 4 or 5 years of age, exhibiting wheezing symptoms anddiagnosed or diagnosable as “wheezy infants”, an established patientcategory of major medical concern and now often identified as incipientor early-phase asthmatics, or as “wheezy-infant syndrome”.

The suitability of a channel activating protease inhibitor such as aprostasin inhibitor for the treatment of a disease mediated byinhibition of a channel activating protease, may be tested bydetermining the inhibitory effect of the channel activating proteaseinhibitor according to the assays described below and following methodsknown in the art.

In accordance with the foregoing, the present invention further providesa method for preventing or treating any of the diseases or disordersdescribed above in a subject in need of such treatment, which methodcomprises administering to said subject a therapeutically effectiveamount of a compound of Formula (1), (2A), (2B), (3A) or (3B), or apharmaceutically acceptable salt thereof. For any of the above uses, therequired dosage will vary depending on the mode of administration, theparticular condition to be treated and the effect desired.

Administration and Pharmaceutical Compositions

In general, compounds of the invention will be administered intherapeutically effective amounts via any of the usual and acceptablemodes known in the art, either singly or in combination with one or moretherapeutic agents.

Channel activating protease inhibitors of the invention are also usefulas co-therapeutic agents for use in combination with other drugsubstances such as anti-inflammatory, bronchodilatory, antihistamine oranti-tussive drug substances, particularly in the treatment of cysticfibrosis or obstructive or inflammatory airways diseases such as thosementioned hereinbefore, for example as potentiators of therapeuticactivity of such drugs or as a means of reducing required dosaging orpotential side effects of such drugs.

The channel activating protease inhibitor may be mixed with the otherdrug substance in a fixed pharmaceutical composition or it may beadministered separately, before, simultaneously with or after the otherdrug substance.

Accordingly, the invention may include a combination of channelactivating protease inhibitor with an anti-inflammatory,bronchodilatory, antihistamine, anti-tussive, antibiotic or DNase drugsubstance, said channel activating protease inhibitor and said drugsubstance being in the same or different pharmaceutical composition.

Suitable anti-inflammatory drugs include steroids, in particularglucocorticosteroids such as budesonide, beclamethasone dipropionate,fluticasone propionate, ciclesonide or mometasone furoate, or steroidsdescribed in international patent application WO 02/88167, WO 02/12266,WO 02/100879, WO 02/00679 (for example, Examples 3, 11, 14, 17, 19, 26,34, 37, 39, 51, 60, 67, 72, 73, 90, 99 and 101), WO 03/35668, WO03/48181, WO 03/62259, WO 03/64445, WO 03/72592, WO 04/39827 and WO04/66920; non-steroidal glucocorticoid receptor agonists, such as thosedescribed in DE 10261874, WO 00/00531, WO 02/10143, WO 03/82280, WO03/82787, WO 03/86294, WO 03/104195, WO 03/101932, WO 04/05229, WO04/18429, WO 04/19935 and WO 04/26248; LTD4 antagonists such asmontelukast and zafirlukast; PDE4 inhibitors such cilomilast (ARIFLO®GlaxoSmithKline), ROFLUMILAST® (Byk Gulden), V-11294A (Napp), BAY19-8004(Bayer), SCH-351591 (Schering-Plough), AROFYLLINE® (AlmirallProdesfarma), PD189659/PD168787 (Parke-Davis), AWD-12-281 (Asta Medica),CDC-801 (Celgene), SelCID™ CC-10004 (Celgene), VM554/UM565 (Vernalis),T-440 (Tanabe), KW-4490 (Kyowa Hakko Kogyo), and those disclosed in WO92/19594, WO 93/19749, WO 93/19750, WO 93/19751, WO 98/18796, WO99/16766, WO 01/13953, WO 03/104204, WO 03/104205, WO 03/39544, WO04/000814, WO 04/000839, WO 04/005258, WO 04/018450, WO 04/018451, WO04/018457, WO 04/018465, WO 04/018431, WO 04/018449, WO 04/018450, WO04/018451, WO 04/018457, WO 04/018465, WO 04/019944, WO 04/019945, WO04/045607 and WO 04/037805; and adenosine A_(2B) receptor antagonistssuch as those described in WO 02/42298, each of which is incorporatedherein in its entirety.

Suitable bronchodilatory drugs include beta-2 adrenoceptor agonists suchas albuterol (salbutamol), metaproterenol, terbutaline, salmeterolfenoterol, procaterol, formoterol, or carmoterol and pharmaceuticallyacceptable salts thereof, and compounds of Formula (1) as described inWO 00/75114 (in free or salt or solvate form), which is incorporatedherein by reference in its entirety, such as a compound of formula:

and pharmaceutically acceptable salts thereof; compounds of Formula (1)of WO 04/16601; as well as compounds of EP 1440966, JP 05025045, WO93/18007, WO 99/64035, US 2002/0055651, WO 01/42193, WO 01/83462, WO02/66422, WO 02/70490, WO 02/76933, WO 03/24439, WO 03/42160, WO03/42164, WO 03/72539, WO 03/91204, WO 03/99764, WO 04/16578, WO04/22547, WO 04/32921, WO 04/33412, WO 04/37768, WO 04/37773, WO04/37807, WO 04/39762, WO 04/39766, WO 04/45618 WO 04/46083 and WO04/80964, each in free or salt or solvate form. Each of thesepublications is incorporated herein in its entirety.

Suitable bronchodilatory drugs also include anticholinergic orantimuscarinic agents, in particular ipratropium bromide, oxitropiumbromide, tiotropium salts and CHF 4226 (Chiesi), glycopyrrolate, andalso those described in EP 424021, U.S. Pat. No. 3,714,357, U.S. Pat.No. 5,171,744, WO 01/04118, WO 02/00652, WO 02/51841, WO 02/53564, WO03/00840, WO 03/33495, WO 03/53966, WO 03/87094, WO 04/018422 and WO04/05285, each of which is incorporated herein in its entirety.

Suitable dual anti-inflammatory and bronchodilatory drugs include dualbeta-2 adrenoceptor agonist/muscarinic antagonists such as thosedisclosed in US 2004/0167167, WO 04/74246 and WO 04/74812, each of whichis incorporated herein in its entirety.

Suitable antihistamine drug substances include cetirizine hydrochloride,acetaminophen, clemastine fumarate, promethazine, loratidine,desloratidine, diphenhydramine and fexofenadine hydrochloride,activastine, astemizole, azelastine, ebastine, epinastine, mizolastineand tefenadine as well as those disclosed in JP 2004107299, WO 03/099807and WO 04/026841, each of which is incorporated herein in its entirety.

Suitable antibiotics include macrolide antibiotics, for exampletobramycin (TOBI™).

Suitable DNase drug substances include dornase alfa (PULMOZYME™), ahighly purified solution of recombinant human deoxyribonuclease I(rhDNase), which selectively cleaves DNA. Dornase alfa is used to treatcystic fibrosis.

Other useful combinations of channel activating protease inhibitors withanti-inflammatory drugs are those with antagonists of chemokinereceptors, e.g. CCR-1, CCR-2, CCR-3, CCR-4, CCR-5, CCR-6, CCR-7, CCR-8,CCR-9 and CCR10, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, particularly CCR-5antagonists such as Schering-Plough antagonists SC-351125, SCH-55700 andSCH-D, Takeda antagonists such asN-[[4-[[[6,7-dihydro-2-(4-methyl-phenyl)-5H-benzo-cyclohepten-8-yl]carbonyl]amino]phenyl]-methyl]tetrahydro-N,N-dimethyl-2H-pyran-4-amin-iumchloride (TAK-770), and CCR-5 antagonists described in U.S. Pat. No.6,166,037, WO 00/66558, WO 00/66559, WO 04/018425 and WO 04/026873, eachof which is incorporated herein in its entirety.

In the treatment of a disease mediated by inhibition of prostasin inaccordance with the invention, a channel activating protease inhibitorof the invention in free form or in pharmaceutically acceptable saltform, may be administered by any appropriate route, for example orally,e.g. in tablet, capsule or liquid form; parenterally, for example in theform of an injectable solution or suspension; intranasally, for examplein the form of an aerosol or other atomisable formulation using anappropriate intranasal delivery device, e.g. a nasal spray such as thoseknown in the art; or by inhalation, such as use with a nebulizer.

The channel activating protease inhibitor may be administered in apharmaceutical composition together with a pharmaceutically acceptablediluent or carrier. Such compositions may be, for example dry powders,tablets, capsules and liquids, but also injection solutions, infusionsolutions or inhalation suspensions, which may be prepared using otherformulating ingredients and techniques known in the art.

The dosage of the channel activating protease inhibitor in free form orin pharmaceutically acceptable salt form may depend on various factors,such as the activity and duration of action of the active ingredient,the severity of the condition to be treated, the mode of administration,the species, sex, ethnic origin, age and weight of the subject and/orits individual condition. In a normal case, the daily dose foradministration, for example oral administration to a warm-bloodedanimal, particularly a human being weighing about 75 kg, is estimated tobe from approximately 0.7 mg to approximately 1400 mg; or in someexamples, from approximately 5 mg to approximately 200 mg. That dose maybe administered in a single dose or in several part doses, for example,from 5 to 200 mg.

When the composition comprises an aerosol formulation, it may contain ahydro-fluoro-alkane (HFA) propellant such as HFA134a or HFA227 or amixture thereof; one or more co-solvents known in the art such asethanol (up to 20% by weight); one or more surfactants such as oleicacid or sorbitan trioleate; and/or one or more bulking agents such aslactose. When the composition comprises a dry powder formulation, it maycontain, for example, the channel activating protease inhibitor having aparticle diameter up to 10 microns, optionally together with a diluentor carrier, such as lactose, of the desired particle size distributionand a compound that helps to protect against product performancedeterioration due to moisture (e.g. magnesium stearate). When thecomposition comprises a nebulised formulation, it may contain, forexample, the channel activating protease inhibitor either dissolved, orsuspended, in a vehicle containing water, a co-solvent such as ethanolor propylene glycol and a stabiliser, which may be a surfactant.

The invention includes (A) a compound of the invention in inhalableform, e.g. in an aerosol or other atomisable composition or in aninhalable particulate, e.g. micronised form; (B) an inhalable medicamentcomprising a compound of the invention in inhalable form; (C) apharmaceutical product comprising a compound of the invention ininhalable form in association with an inhalation device; and (D) aninhalation device containing a compound of the invention in inhalableform.

Processes for Making Compounds of the Invention

The present invention also includes processes for the preparation ofcompounds of the invention. In the reactions described, reactivefunctional groups, where desired in the final product (e.g., hydroxy,amino, imino, thio or carboxy groups), may be protected using protectinggroups known in the art, to avoid their unwanted participation in thereactions. Conventional protecting groups may be used in accordance withstandard practice, for example, see T. W. Greene and P. G. M. Wuts in“Protective Groups in Organic Chemistry”, John Wiley and Sons, 1991.

In one embodiment, compounds of the invention may be prepared followingthe Reaction scheme I below:

wherein R^(1′) is (CR₂)_(n)—X_(i)—R¹;

R^(2′) is (CR₂)_(k)R²;

R^(4′) is (CR₂)_(m)—R⁴;

R¹, R², R⁴, X, Y, Z, i, k, m and n are as defined in Formula (1);

R⁷ and R⁸ are alkyl protecting groups (for example, methyl, ethyl,t-butyl or benzyl and the like).

In the above Reaction Scheme I, the intermediate compound II may besynthesized by reacting intermediate compound Iaa with an alkyl reagentof the type R²—X where X is a leaving group, in the presence of asuitable base and a suitable organic solvent. Examples of leaving groupsin alkyl reagents R²—X include but are not limited to halides such aschlorides and bromides, or a tosylate, mesylate, or besylate leavinggroups, and the like. Intermediate compound III may be prepared byreacting intermediate compound II with diazomethane or(trimethylsilyl)diazomethane in the presence of a suitable organicsolvent. These reactions may proceed in a temperature range of about 0°C. to about 60° C., and may take up to about 24 hours to complete.

Alternatively, intermediate compound III may be synthesized by reactingintermediate compound II with an alkyl reagent of the type R⁸—X where Xis a leaving group as previously described above, in the presence of asuitable base and a suitable organic solvent. The reaction may proceedin a temperature range of about 0° C. to about 80° C., and may take upto about 24 hours to complete.

Alternatively, intermediate compound III may be synthesized by reactingintermediate compound II with an alcohol of the type R⁸—OH with asuitable peptide coupling reagent and a suitable base in the presence ofa suitable solvent. Suitable bases for this reaction include but are notlimited to triethylamine, DIEA, pyridine, 2,4,6-collidine, and othersuitable bases within the knowledge of those skilled in the art. Thereaction may proceed in a temperature range of about 0° C. to about 40°C., and may take up to about 24 hours to complete.

In the above Reaction Scheme I, intermediate compound IV may besynthesized by removing the carbamate protecting group (e.g., where R⁷is t-butyl) from intermediate compound III with a suitable acid, andoptionally in the presence of a suitable organic solvent. Suitable acidsinclude but are not limited to TFA, p-TsOH, TfOH, HCl, HBr, HF, HBF₄,and other suitable acids within the knowledge of those skilled in theart. The reaction may proceed in a temperature range of about −20° C. toabout 40° C., and may take up to about 24 hours to complete.

Alternatively, intermediate IV may be synthesized by removing thecarbamate protecting group from intermediate compound III (e.g., whereR⁷ is benzyl or any benzylic derivative) with hydrogen gas in thepresence of a suitable catalyst and a suitable solvent or water.Examples of suitable catalysts include but are not limited to Pd/C, Pt,PtO₂, Pt/C, Rh/C, and other suitable catalysts within the knowledge ofthose skilled in the art. The reaction may proceed in a temperaturerange of about 0° C. to about 80° C., with hydrogen pressures of about15 psi to about 80 psi, and may take up to about 48 hours to complete.

Intermediate compound VI may be synthesized by reacting intermediatecompounds IV and V in the presence of a peptide coupling reagent and asuitable base (Et₃N, DIEA, pyridine, 2,4,6-collidine, and the like) inthe presence of a suitable organic solvent. The reaction may proceed ina temperature range of about 0° C. to about 40° C., and may take up toabout 24 hours to complete. Intermediate compound VII may be synthesizedby reacting intermediate compound VI with a suitable base (for exampleLiOH, NaOH, KOH, K₂CO₃, NaCO₃, CsCO₃, and the like) in the presence of asuitable organic solvent or water. The reaction may proceed in atemperature range of about 0° C. to about 40° C., and may take up toabout 24 hours to complete. Intermediate compound of IX may besynthesized by reacting intermediate compounds VII and VIII in thepresence of a suitable peptide coupling reagent and a suitable base(Et₃N, DIEA, pyridine, 2,4,6-collidine, and the like) in the presence ofa suitable organic solvent. The reaction may proceed in a temperaturerange of about 0° C. to about 40° C., and may take up to about 24 hoursto complete.

The final compound X may be synthesized by reacting intermediatecompound IX with a suitable oxidant in the presence of a suitableorganic solvent or water. Suitable oxidants include but are not limitedto for Dess-Martin periodinane, 2-iodobenzoic acid with oxone, TEMPOwith trichlorisocyanuric acid, TEMPO with NaOCl, DMSO with oxalylchloride, pyridinium chlorochromate, MnO₂, CrO₂, and other suitableoxidants within the knowledge of those skilled in the art. The reactionmay proceed in a temperature range of about 0° C. to about 40° C., andmay take up to about 24 hours to complete.

In another embodiment, compounds of the invention may be preparedfollowing the Reaction scheme II below, and following conditions forsimilar reactions as described in Reaction Scheme I:

wherein R^(1′), R^(2′), R^(4′), Y and Z are as described in ReactionScheme I.

In the above Reaction Scheme II, intermediate compound II may besynthesized by reacting intermediate compound Iaa with an alkyl reagentof the type R₂—X where X is a leaving group, in the presence of a baseand solvent. Intermediate compound IV may be synthesized by reactingintermediate compounds II and III with a peptide coupling reagent and abase in a solvent. Intermediate compound V may be synthesized byremoving the carbamate protecting group from intermediate compound IV(e.g., where R⁷ is t-butyl), by reacting intermediate compound IV withan acid, and optionally in a solvent. Alternatively, intermediatecompound V may be synthesized by removing the carbamate protecting groupfrom intermediate compound IV (where R⁷ is benzyl or any benzylicderivative), with hydrogen gas in the presence of a catalyst in anorganic solvent or water. Intermediate compound VII may be synthesizedby reacting intermediate compounds V and VI with a peptide couplingreagent and a base in solvent. The final compound VIII may besynthesized by reacting intermediate compound VII with an oxidant in anorganic solvent or water.

In yet another embodiment, compounds of formula (2) may be preparedfollowing the Reaction Scheme III below, and following conditions forsimilar reactions as described in Reaction Scheme I:

wherein R^(1′), R^(4′), R⁶, R⁷, Y and Z are as defined in Formula (1),and X is an activated leaving group such as —OPhNO₂, or —Cl.

In the above Reaction Scheme III, intermediate compound II may besynthesized by reacting intermediate compound Iaa withtrans-4-hydroxyproline methyl ester and a peptide coupling reagent inthe presence of a solvent and base. The reaction may proceed at roomtemperature, and may take up to about 24 hours to complete.

Intermediate compound III may be synthesized by reacting intermediatecompound II with a suitable reactive chemical intermediate (for example,phenyl chloroformate, 4-nitrophenyl chloroformate, pentafluorophenylchloroformate and the like) and a suitable base in a suitable solvent.The reaction may proceed in a temperature range of about 0° C. to about60° C., and may take up to about 24 hours to complete.

Intermediate compound IV may be synthesized by reacting intermediatecompound III with a suitable primary or secondary amine in the presenceof a suitable solvent. The reaction may proceed in a temperature rangeof about 0° C. to about 60° C., and may take up to about 24 hours tocomplete. Intermediate compound V may be synthesized by reactingintermediate compound IV with a suitable base (for example LiOH, NaOH,KOH and the like) in a suitable organic solvent with or without water.The reaction may proceed in a temperature range of about 0° C. to about40° C., and may take up to about 24 hours to complete. Intermediatecompound VII may be synthesized by reacting intermediate compound V withthe intermediate amine VI in the presence of a peptide coupling reagentand a base in solvent. The final compound VIII may be synthesized byreacting intermediate compound VII with a suitable oxidant in a solvent.

Suitable peptide coupling reagents for use in the reactions described inReaction schemes I, II and III include but are not limited to DCC, DIC,HATU, BOP, PyBOP, EDC, and other coupling reagents within the knowledgeof those skilled in the art.

Suitable bases for use in the reactions described in Reaction schemes I,II and III include but are not limited to hydroxides such as NaOH, KOH,or LiOH; carbonates such as K₂CO₃ or CsCO₃; hydrides such as NaH or KH,and the like. Other suitable bases are amines, DIEA, pyridine,2,4,6-collidine, and other suitable bases within the knowledge of thoseskilled in the art.

Suitable organic solvents for use in the reactions described in Reactionschemes I, II and III include but are not limited to DMSO, THF, DMF,DMAc, acetonitrile, acetone, 2-propanone, butanone, HMPA, NMP,dichloromethane, chloroform, 1,2-dichloroethane, diethyl ether,methanol, ethanol, t-butanol, isopropanol, propanol, n-butanol,cyclohexanol, acetonitrile, dioxane, MTBE, benzene, toluene, andmixtures thereof, and other suitable solvents within the knowledge ofthose skilled in the art.

Additional Processes for Making Compounds of the Invention

A compound of the invention may be prepared as a pharmaceuticallyacceptable acid addition salt by reacting the free base form of thecompound with a pharmaceutically acceptable inorganic or organic acid.Alternatively, a pharmaceutically acceptable base addition salt of acompound of the invention may be prepared by reacting the free acid formof the compound with a pharmaceutically acceptable inorganic or organicbase.

Alternatively, the salt forms of the compounds of the invention may beprepared using salts of the starting materials or intermediates.

The free acid or free base forms of the compounds of the invention maybe prepared from the corresponding base addition salt or acid additionsalt from, respectively. For example, a compound of the invention in anacid addition salt form may be converted to the corresponding free baseby treating with a suitable base (e.g., ammonium hydroxide solution,sodium hydroxide, and the like). A compound of the invention in a baseaddition salt form may be converted to the corresponding free acid bytreating with a suitable acid (e.g., hydrochloric acid, etc.).

Compounds of the invention in unoxidized form may be prepared fromN-oxides of compounds of the invention by treating with a reducing agent(e.g., sulfur, sulfur dioxide, triphenyl phosphine, lithium borohydride,sodium borohydride, phosphorus trichloride, tribromide, or the like) ina suitable inert organic solvent (e.g. acetonitrile, ethanol, aqueousdioxane, or the like) at 0 to 80° C.

Prodrug derivatives of the compounds of the invention may be prepared bymethods known to those of ordinary skill in the art (e.g., for furtherdetails, see Saulnier et al., (1994), Bioorganic and Medicinal ChemistryLetters, Vol. 4, p. 1985). For example, appropriate prodrugs may beprepared by reacting a non-derivatized compound of the invention with asuitable carbamylating agent (e.g., 1,1-acyloxyalkylcarbanochloridate,para-nitrophenyl carbonate, or the like).

Protected derivatives of the compounds of the invention may be made bymeans known to those of ordinary skill in the art. A detaileddescription of techniques applicable to the creation of protectinggroups and their removal may be found in T. W. Greene, “ProtectingGroups in Organic Chemistry”, 3^(rd) edition, John Wiley and Sons, Inc.,1999.

Compounds of the present invention may be conveniently prepared, orformed during the process of the invention, as solvates (e.g.,hydrates). Hydrates of compounds of the present invention may beconveniently prepared by recrystallization from an aqueous/organicsolvent mixture, using organic solvents such as dioxin, tetrahydrofuranor methanol.

Compounds of the invention may be prepared as their individualstereoisomers by reacting a racemic mixture of the compound with anoptically active resolving agent to form a pair of diastereoisomericcompounds, separating the diastereomers and recovering the opticallypure enantiomers. Resolution of enantiomers may be carried out usingcovalent diastereomeric derivatives of the compounds of the invention,or by using dissociable complexes (e.g., crystalline diastereomericsalts). Diastereomers have distinct physical properties (e.g., meltingpoints, boiling points, solubilities, reactivity, etc.), and may bereadily separated by taking advantage of these dissimilarities. Thediastereomers may be separated by chromatography, or byseparation/resolution techniques based upon differences in solubility.The optically pure enantiomer is then recovered, along with theresolving agent, by any practical means that would not result inracemization. A more detailed description of the techniques applicableto the resolution of stereoisomers of compounds from their racemicmixture may be found in Jean Jacques, Andre Collet, Samuel H. Wilen,“Enantiomers, Racemates and Resolutions”, John Wiley And Sons, Inc.,1981.

In summary, the compounds of Formula (1) may be made by a process, whichinvolves:

(a) that of Reaction Scheme I, II or III;

(b) optionally converting a compound of the invention into apharmaceutically acceptable salt;

(c) optionally converting a salt form of a compound of the invention toa non-salt form;

(d) optionally converting an unoxidized form of a compound of theinvention into a pharmaceutically acceptable N-oxide;

(e) optionally converting an N-oxide form of a compound of the inventionto its unoxidized form;

(f) optionally resolving an individual isomer of a compound of theinvention from a mixture of isomers;

(g) optionally converting a non-derivatized compound of the inventioninto a pharmaceutically acceptable prodrug derivative; and

(h) optionally converting a prodrug derivative of a compound of theinvention to its non-derivatized form.

Insofar as the production of the starting materials is not particularlydescribed, the compounds are known or may be prepared analogously tomethods known in the art or as disclosed in the Examples hereinafter.One of skill in the art will appreciate that the above transformationsare only representative of methods for preparation of the compounds ofthe present invention, and that other well known methods may similarlybe used. The present invention is further exemplified, but not limited,by the following intermediates (Reference compounds) and Examples thatillustrate the preparation of the compounds of the invention.

In the synthetic methodologies below, the following common abbreviationsknown in the art are used: DCM (dichloromethane); THF (tetrahydrofuran);and DIEA (diisopropylethylamine).

In the above Scheme 1, the reagents and conditions are: (a) iso-BuOCOCl,Et₃N, THF; NaBH₄, H₂O. (b) Dess-Martin periodinane, CH₂Cl₂; (c)iso-PrMgCl, benzoxazole, THF, −20° C., 30 min, then 1-C, −20° C. to rt.(d) H₂ (40 psi), EtOH, Pd/C 10%, rt, 18 h.

1-B: The crude starting material, Z-Lys(Boc)OH (320 g, 842 mmol) isdissolved in THF (2500 mL) and the solution is cooled to −10° C.followed by the addition of triethylamine (115.2 mL, 1.0 eq) anddropwise addition of iso-butylchloroformate (118.7 mL, 1.1 eq). Theresulting suspension is stirred for 2 h at 0° C. The reaction mixture isfiltered and cooled to −10° C. NaBH₄ (64.6 g, 2.1 eq) is dissolved inwater (500 mL) at 0° C. and the solution is added portionwise to the THFsolution (heavy CO₂ evolution). The reaction mixture is allowed to warmto room temperature and stirred for one hour. The reaction mixture isacidified with 1N HCl solution, and the aqueous phase is extractedseveral times with EtOAc. The combined organic layers are washed withwater, saturated aqueous NaHCO₃ solution and brine; dried on MgSO₄; andthe solvent is removed in vacuo. The product is purified by flash columnchromatography (hexanes/ethyl acetate) to afford the desired product asa white foam.

1-C: The alcohol (200 g, 545.8 mmol) is dissolved in DCM (2000 mL) andcooled to 0° C. A solution of the Dess-Martin reagent (231 g, 1.0 eq) inDCM (2000 mL) is added portionwise. The suspension is allowed to warm toroom temperature and stirred until complete conversion (1-4 hours). A1:1 mixture of saturated aqueous NaHCO₃ solution and a 1M Na₂S₂O₃solution is added and the resulting biphasic system is stirredvigorously for 20 minutes. The organic layer is separated and theaqueous layer is extracted one time with DCM. The combined organiclayers are distilled in vacuo and the resulting oil is taken up in EtOAcand washed six times with the NaHCO₃/Na₂S₂O₃ mixture, water and brine;dried on MgSO₄ and the solvent is removed in vacuo to give the crudealdehyde as a yellowish oil. The material is directly used in the nextstep without further purification.

1-D: To a solution of isopropyl-magnesium chloride (1.67 eq. vsaldehyde, 390 mL of a 2M-THF solution from Sigma-Aldrich) in THF (1500mL) is added benzoxazole (92.8 g, 1.67 eq) in THF (1000 mL) at −20° C.The reaction mixture is stirred at −20° C. for 30 minutes (color change:deep red) and a solution of the aldehyde (170 g, 466 mmol) in THF (1500mL) is added slowly under temperature control at −20° C. to −15° C. Thereaction mixture is allowed to warm to room temperature and stirreduntil completion. The reaction mixture is quenched with saturatedaqueous NH₄Cl solution and the solvent is removed in vacuo. The aqueousphase is extracted three times with EtOAc; the combined organic layersare excessively washed with 1N HCl solution, water, brine, dried onMgSO₄, and the solvent is removed in vacuo to give the crude benzoxazoleas a deep red oil. Purification on silica with EtOAc/hexanes (1:5 to1:1) gave the benzoxazole as a yellow solid.

1-E: A solution of Compound 5 (25.0 g, 51.7 mmol) is dissolved inethanol (150 mL). Pd/C (10%, wet, Degussa type) is added, and the flaskis placed on a Parr shaker overnight and subjected to hydrogen gas at 40psi. The catalyst is filtered through Celite, and solvent is removed invacuo. The crude material is purified by flash chromatography usingfirst a gradient of hexanes/EtOAc to remove less polar and coloredimpurities, then followed by a gradient of DCM/MeOH to elute the desiredcompound 5. The solvent is removed in vacuo, and the compound istriturated several times in ether to afford the desired referencecompound 1 as a white powder. ¹H-NMR (DMSO-d6, 400 MHz) δ 7.73-7.70 (2H,m), 7.40-7.34 (2H, m), 6.78-6.73 (1H, m), 4.55-4.51 (1H, m), 3.05-3.01(1H, m), 2.92-2.83 (2H, m), 1.48-1.18 (14H, m). LCMS: 350.5 (M+H)⁺.

In the above Scheme 2, the reagents and conditions are: a) Cbz-OSu,Et₃N, THF, H₂O, rt, 18 h, (b) i. iso-BuOCOCl, Et₃N, THF; ii. NaBH₄, H₂O;(c) Dess-Martin periodinane, CH₂Cl₂; (d) iso-PrMgCl, benzoxazole, THF,−20° C., 30 min, then 2-D, −20° C. to rt; (e) Indium, NH₄CT, EtOH,reflux, 5 h; (f)N,N-Bis(tert-butoxycarbonyl)-1H-pyrazole-1-carboxamidine, DIEA, MeOH;(g) H₂, (40 psi), 10% Pd/C, EtOH.

2-B: L-Nitrophenylalanine hydrochloride (4.45 g, 18.0 mmol) andN-(Benzyloxycarbonyloxy)succinimide (Cbz-OSu) (4.49 g, 18.0 mmol) areadded to a round bottomed flask containing THF (60 mL) and water (20mL). The mixture is stirred at room temperature and Et₃N (10.1 mL, 72.0mmol) is added, and the reaction is stirred overnight at roomtemperature. The clear solution is diluted with EtOAc (200 mL), andwashed with 1N HCl (3×100 mL) and brine (1×100 mL) and dried with MgSO₄.Solvent is evaporated in vacuo to afford intermediate 2-B as a whitesolid.

2-C to 2-E: These intermediates are prepared following methods analogousto those described for preparing intermediates 1-B to 1-D of Referencecompound 1, respectively.

2-F: The nitrophenyl analog 5-E (1.85 g, 4.15 mmol) is dissolved in EtOH(50 mL) and heated to reflux. Saturated aqueous NH₄Cl (5 mL) is added,followed by powdered indium (3.2 g, 27.9 mmol). The reaction mixture isstirred at reflux for 5 h, cooled to room temperature, and the solventis removed in vacuo. The crude material is suspended in EtOAc (100 mL),washed with saturated NaHCO₃ (3×100), dried with MgSO₄, and filteredthrough Celite. The solvent is removed in vacuo to afford the aniline2-F as an off-white waxy solid.

2-G: Aniline 2-F (1.52 g, 3.67 mmol) is dissolved in MeOH (10 mL). DIEA(0.7 mL, 4.4 mmol) andN,N-Bis(tert-butoxycarbonyl)-1H-pyrazole-1-carboxamidine (1.37 g, 4.4mmol) are added, and the reaction mixture is stirred at roomtemperature. After 4 h, another 0.5 equiv ofN,N-Bis(tert-butoxycarbonyl)-1H-pyrazole-1-carboxamidine (0.685 g, 2.2mmol) is added and the reaction is then stirred overnight at roomtemperature. EtOAc (100 mL) is added, and the organic layer is washedwith water, brine, dried on MgSO₄. The solvent is removed in vacuo, andthe crude material is purified by silica gel chromatography withEtOAc/hexanes (0 to 100% gradient) to afford the desired product 2-G asan oil.

2-H: This compound is prepared from 2-G following methods analogous tothose described for the preparation of intermediate 1-E for Referencecompound 1.

In the above Scheme 3, the reagents and conditions are: (a)HN(OMe)Me.HCl, BOP, Et₃N, DMF, 0° C. to rt; (b) n-BuLi (2.5 M inhexanes), benzothiazole, THF, −78° C., then 3-B, THF, −70° C. to rt; (c)NaBH₄, MeOH; (d) p-TsOH, CH₂Cl₂, 6 h.

3-B: BOP (50 g, 112 mmol) is added in one portion to a stirring solutionof 3-A (49.92 g, 102.6 mmol), N,O-dimethylhydroxylamine hydrochloride(30.4 g, 224 mmol) and triethylamine (88 ml, 616 mmol) in dry DMF (200mL) under argon at 0° C. The reaction mixture is allowed to slowly warmto room temperature over 2 h, filtered through diatomaceous earth, andconcentrated in vacuo. The residue is dissolved in ethyl acetate; washedwith H₂O, 1 M aqueous KHSO₄, saturated aqueous NaHCO₃, and brine; dried,and concentrated in vacuo. The residue is purified by silica gel columnchromatography to give compound 3-B.

3-C: n-Butyllithium (2.5 M in hexanes, 272.2 ml, 681.4 mmol) is addeddropwise at −78° C. under argon to a stirring solution of benzothiazole(115.72 g, 850.7 mmol) in dry THF (1660 mL) at a rate that kept thereaction temperature below −64° C. Upon completion of addition, thereaction mixture is stirred for 30 min at −70° C., and a solution ofcompound 3-B (45 g, 85.7 mmol) in dry THF (300 ml) is added at a ratethat maintained the reaction temperature below −70° C. The reaction isstirred for 15 min, quenched with saturated aqueous NH₄Cl, and stirredfor 16 h at room temperature. The resulting organic layer is separated;diluted with ethyl acetate; washed with water and brine; dried andconcentrated in vacuo; and purified by silica gel chromatography to givecompound 3-C.

3-D: To a solution of 3-C (33.7 g, 55.82 mmol) in MeOH (407 ml) at 0° C.is added NaBH₄ (9.98 g). The reaction mixture is slowly warmed to roomtemperature over 1 h, then heated to 45° C. for 1 h, and then cooledback to room temperature. The reaction is quenched with acetone (60 mL),and concentrated in vacuo. The residue is dissolved in ethyl acetate,washed with brine, and dried over MgSO₄. The crude material is purifiedby silica gel chromatography to afford product 3-D.

3-E: p-TsOH is added to a stirring solution of compound 3-D (28.2 g) inCH₂Cl₂ (300 mL) at room temperature until the solution is saturated. Thereaction is stirred at room temperature for 6 h. Water is added, and theorganic layer is extracted with EtOAc; washed with 1:1 mixture (V/V) ofbrine and 10% aqueous Na₂CO₃; dried over Na₂SO₄; and purified by silicagel chromatography to give the product 3-E.

In the above Scheme 4, the reagents and conditions are: (a) i. NaN₃,Tf₂O, H₂O, CH₂Cl₂, 0° C.; ii. Cbz-Orn-OH, K₂CO₃, H₂O, CuSO₄, then TfN₃,MeOH; (b) i. iso-BuOCOCl, Et₃N, THF; ii. NaBH₄, H₂O; (c) Dess-Martinperiodinane, CH₂Cl₂; (d) iso-PrMgCl, benzoxazole, THF, −20° C., 30 min,then 4-D, −20° C. to rt; (e) PMe₃, THF, H₂O (f)N,N-Bis(tert-butoxycarbonyl)-1H-pyrazole-1-carboxamidine, MeOH; (g) H₂,(40 psi), 10% Pd/C, EtOH.

4-B: Preparation of the triflic azide (TfN₃) solution: Sodium azide (61g, 938.8 mmol, 10 eq) is dissolved in water (150 mL), and the solutionis cooled to 0° C. Dichloromethane (250 mL) is added and the biphasicsystem is stirred vigorously while freshly distilledtrifluoromethansulfonylanhydride (Tf₂O) is added over a period of 30min. The reaction mixture is stirred vigorously for an additional twohours at 0° C. The phases are separated, and the aqueous layer isextracted twice with dichloromethane (each 100 mL). The combined organiclayers are washed twice with saturated aqueous NaHCO₃ solution (each 100ml). This solution is kept and used as is.

Z-Orn-OH (25 g, 93.88 mmol) is dissolved in water (250 mL) followed bythe addition of potassium carbonate (18.16 g, 131.4 mmol, 1.4 eq) andCuSO₄ (1 mol %, 250 mg). The TfN₃ solution is added at once to thereaction mixture at room temperature. After complete addition (biphasicmixture), methanol is added until the reaction mixture became monophasic(approximately 850 mL), and the resulting reaction solution is stirredfor 24 to 48 h. The solvents are evaporated, and the blue aqueous phaseis acidified with 1 M NaHSO₄ (color disappeared). The aqueous phase isextracted with ethyl acetate (3×400 mL), and the combined organic layersare washed with water and brine; dried on MgSO₄; and the solvent isremoved in vacuo to give crude azide which is purified over silica gelchromatography with EtOAc/hexanes (1:9 to 1:1) as eluent to give theazide 4-B as a yellowish oil.

4-C to 4-E: Intermediates 4-C to 4-E are prepared following methodsanalogous to those described for preparing the intermediates ofReference compound 1.

4-F: Azide 4-E (6.275 g, 15.9 mmol) is dissolved in a mixture ofTHF/water (10:1, 100 mL). Trimethylphosphine (2.0 eq) is added slowly atroom temperature until the complete conversion is observed by LCMS. Thesolvent is removed and the residue is taken up in EtOAc. The organiclayer is washed with water, brine dried on MgSO₄, and the solvent isremoved in vacuo to give the crude amine 4-F which is used in the nextstep without further purification.

4-G: Amine 4-F (5.608 g, 15.2 mmol) is dissolved in MeOH (100 mL) andN,N-Bis (tert-butoxycarbonyl)-1H-pyrazole-1-carboxamidine (5.189 g, 1.1eq) is added, and the reaction mixture is stirred for two hours at roomtemperature. The solvent is removed in vacuo, and the residue is takenup in EtOAc. The organic layer is washed with water and brine; dried onMgSO₄, and the solvent is removed in vacuo to give the crudebenzoxazole-derivative. Purification on silica with EtOAc/hexanes (0 to100% EtOAc) affords the final benzoxazole derivative as an oil.

4-H: This compound is prepared from 4-G following methods analogous tothose described for the preparation of intermediate 1-E for Referencecompound 1.

In the above Scheme 5, the reagents and conditions are: a) Cbz-OSu,Et₃N, THF, H₂O, rt, 18 h; (b) i. iso-BuOCOCl, Et₃N, THF; ii. NaBH₄, H₂O;(c) Dess-Martin periodinane, CH₂Cl₂; (d) iso-PrMgCl, benzoxazole, THF,−20° C., 30 min, then 5-D, −20° C. to rt; (e) NaBH₄, NiCl₂, Boc₂O, MeOH,0° C., (f) H₂, (40 psi), 10% Pd/C, EtOH.

5-B to 5-E: This compound is prepared from L-4-cyanophenylalanine (5-A)following methods analogous to those described for intermediate 2-B forReference compound 2. Intermediates 5-C to 5-E are prepared followingmethods analogous to those described for preparing intermediates 1-B to1-D of Reference Compound 1, respectively.

5-F: Nitrile 5-E (3.68 g, 8.64 mmol) is dissolved in methanol (60 mL),and cooled to 0° C. Boc₂O (3.77 g, 2 eq) is added, followed by NiCl₂(210 mg, 0.1 eq). The mixture is stirred, then NaBH₄ (2.29 g, 7.0 eq) isadded slowly in small portions. The mixture is warmed to roomtemperature and stirred for 1 hr. Diethylene triamine (0.94 mL, 1 eq) isadded. The mixture stirred for an additional 30 min, and the solvent isevaporated in vacuo. The residue is dissolved in EtOAc, washed withsaturated NaHCO₃ (2×100 mL) twice, then dried with MgSO₄ and the solventis evaporated in vacuo. The crude material is purified by silica gelflash chromatography with 0-100% gradient of EtOAc and hexanes toprovide the title compound as an oil.

5-G: This compound is prepared from 5-F following methods analogous tothose described for intermediate 1-E of Reference compound 1.

In the above Scheme 6, the reagents and conditions are: (a) HONH₂ 50%aq., EtOH, 40° C., 16 h; (b) SmI₂, THF, rt; (c) (e) H₂, (40 psi), 10%Pd/C, EtOH.

6-A: The nitrile intermediate 5-E from Reference compound 5 (1.92 g,4.50 mmol) is dissolved in ethanol, and aqueous HONH₂ (50% aq., 1.1 mL,18.0 mmol) is added. The reaction mixture is heated to 40° C. andstirred for 16 h. The solvent is evaporated, and the residue dissolvedin EtOAc (100 mL) and washed with water (2×50 mL). The organic layer isdried (MgSO₄) and evaporated in vacuo to give the product as clear oilwhich is used in the next step without further purification.

6-B: The hydroxyamidine 6-A (2.05 g, 4.50 mmol) is dissolved in aminimum volume of THF (˜2 mL), and SmI₂ 1M in THF (100 ml, 10 mmol) isadded, and the solution is stirred for 1 h. An additional aliquot ofSmI₂ (1M in THF solution) is added if the solution lost its blue color,and stirred for another hour. The solvent is evaporated, and the residuedissolved in EtOAc (100 mL) and washed with saturated aqueous NaHCO₃(2×50 mL) and brine (50 mL). The organic layer is dried (MgSO₄) andevaporated in vacuo to give a clear oil which is used in the next stepwithout further purification.

6-C: The amidine 6-B (2.10 g, 4.50 mmol) is dissolved in THF (50 mL),and Boc₂O (1.96 g, 9.00 mmol) and Et₃N (1.37 mL, 13.5 mmol) are added.The reaction is stirred at room temperature for 18 h, then the solventis removed in vacuo. The residue is dissolved in EtOAc (100 mL), andwashed with saturated aqueous NaHCO₃ (2×50 mL) and brine (50 mL). Theorganic layer is dried (MgSO₄) and evaporated in vacuo. The cruderesidue is purified by flash chromatography using a gradient of 0-100%EtOAc and hexanes to give a white foam.

6-D: This compound is prepared from 6-C following methods analogous tothose described for the preparation of intermediate 1-E for Referencecompound 1 in Scheme 1.

In the above Scheme 7, the reagents and conditions are: a) i.iso-BuOCOCl, Et₃N, THF; ii. NaBH₄, H₂O; (b) trichloroisocyanuric acid,TEMPO, 0° C. to rt, CH₂Cl₂, (c) Cbz-α-phosphonoglycine trimethyl ester,DBU, DCM; (d) H₂ (60 psi), (S,S)-Me-BPE-Rh(COD)⁺OTf⁻, MeOH, 96 h; (e)LiOH, Dioxane, water; (f) i. iso-BuOCOCl, Et₃N, THF; ii. NaBH₄, H₂O; (g)Dess-Martin periodinane, CH₂Cl₂; (h) iso-PrMgCl, benzoxazole, THF, −20°C., 30 min, then 7-G, −20° C. to rt; (i) H₂, (40 psi), 10% Pd/C, EtOH.

7-A: This compound is prepared fromtrans-4-(tert-Butoxycarbonylaminomethyl)cyclohexanecarboxylic acid (7-A)following methods analogous to those described for the preparation ofintermediate 1-B for Reference compound 1 in Scheme 1.

7-B: Alcohol 7-A (4.14 g, 17.0 mmol) is dissolved in CH₂Cl₂ (35 mL), andthe solution is cooled to 0° C. Trichloroisocyanuric acid (4.15 g, 17.8mmol) is added, followed by TEMPO (28 mg, 0.17 mol). The reaction isthen warmed to room temperature and stirred for a further 15 min at roomtemperature. A precipitate formed, and the reaction mixture is filteredthrough Celite and washed with CH₂Cl₂. The combined CH₂Cl₂ solution(˜100 mL) is washed with saturated aqueous NaHCO₃ (2×50 mL), 1M HCl(2×50 mL), and brine (50 mL); dried (MgSO₄) and the solvent isevaporated to give the intermediate which is used without furtherpurification.

7-C: N-Benzyloxycarbonyl-α-phosphonoglycine trimethyl ester (5.63 g, 17mmol) is dissolved in CH₂Cl₂ (35 mL) and DBU (5.1 mL, 34 mmol) is added,and the solution is stirred for 20 min. The aldehyde 7-B (4.09 g, 17.0mmol) is added dropwise as a solution in CH₂Cl₂ (10 mL). The reaction isstirred overnight, then the solvent is evaporated and the residuedissolved in EtOAc (100 mL) and washed with 1M NaHSO₄ (2×50 mL) andbrine, then dried (MgSO₄) and evaporated in vacuo. The crude material ispurified by flash chromatography using a gradient of 0-100%EtOAc/Hexanes to afford the desired product as a white solid.

7-D: The olefin 7-C (2.04 g, 4.56 mmol) is dissolved in MeOH (100 mL)and the solution is degassed prior to addition of the catalyst,(−)-1,2-Bis-((2S,3S)-2,5-dimethylphospholano)ethane(cyclooctadiene)-rhodium(I)-trifluoro-methanesulfonate (28 mg, 1 mol %). The reaction mixture is placed in a Parrshaker and shaken under 60 psi of H₂ for 4 days. The solvent is thenevaporated in vacuo and the crude material purified by flashchromatography using a gradient of 0-100% EtOAc/Hexanes to afford thedesired product as a white solid.

7-E: The methyl ester 7-D (1.81 g, 4.04 mmol) is dissolved in dioxane(50 mL) and stirred at 0° C. LiOH (203 mg, 4.84 mmol) dissolved in water(10 mL) is added dropwise, and the solution is then warmed to roomtemperature. After the starting material had disappeared (by LCMS), thesolvent is evaporated, and the crude material dissolved in EtOAc (100mL); washed with 1 N NaHSO₄ (2×50 mL) and brine (50 mL); and dried(MgSO₄). The solvent is removed in vacuo and the product is useddirectly in the next step without further purification.

7-F to 7-H: Intermediates 7-F to 7-H are prepared following methodsanalogous to those described for preparing the intermediates ofReference compound 1.

7-I: This compound is prepared from 7-H following methods analogous tothose described for the preparation of intermediate 1-E for Referencecompound 1. MS m/z 404.2 (M+1); 1H NMR (CDCl3, 400 MHz) d 7.66-7.64 (1H,m), 7.49-7.47 (1H, m), 7.31-7.26 (2H, m), 4.85-4.64 (1H, m), 3.44-3.16(1H, m), 2.96-2.88 (2H, m), 1.80-1.55 (4H, m), 1.39-1.14 (13H, m),0.89-0.72 (4H, m).

Reference compound 8 is prepared starting from 3-nitrophenylalaninefollowing methods analogous to those described for the preparation ofReference compound 2.

Reference compound 9 is prepared starting from 3-cyanophenylalaninefollowing methods analogous to those described for the preparation ofReference compound 5.

Reference compound 10 is prepared starting from 3-cyanophenylalaninefollowing methods analogous to those described for the preparation ofReference compound 6.

Reference compound 11 is prepared starting fromtrans-4-(tert-butoxycarbonylamino) cyclohexanecarboxylic acid followingmethods analogous to those described for the preparation of Referencecompound 7.

D-Homophenylalanine ethyl ester hydrochloride (2.43 g, 10.0 mmol) andEt₃N (1.4 mL, 20.0 mmol) are dissolved in DMF (20 mL) and stirred atroom temperature. Phenyl isocyanate (1.55 g, 13.0 mmol) is addeddropwise and the reaction mixture is stirred overnight. The reaction isdiluted with EtOAc (200 mL), and washed with water (100 mL), 1N HCl(2×100 mL), saturated aqueous NaHCO₃ (2×100 mL), brine (100 mL), anddried with MgSO₄. The solvent is evaporated to dryness, and the residueis dissolved in dioxane (20 mL). LiOH.H₂O (630 mg, 15.0 mmol) dissolvedin water (15 mL) is added, and the reaction mixture stirred at roomtemperature until the ethyl ester had disappeared (by TLC and LCMS). Thesolvent is removed in vacuo and the crude material is partitioned withEtOAc (100 mL) and 1N HCl (100 mL). The aqueous layer is extracted withEtOAc (2×50 mL) and the combined organic phases are washed with 1MNaHSO₄ (2×50 mL) and brine (100 mL), and dried with MgSO₄. The solventis evaporated and the crude material purified by flash chromatography(EtOAc:Hexanes gradient) to afford Reference compound 12 as a whitepowder.

N-BOC-cis-4-hydroxy-D-proline (7.83 g, 34 mmol) is dissolved in DMF (100mL) and cooled to 0° C. NaH 60% in mineral oil (3.0 g, 74.8 mmol) isadded portionwise and stirred for 30 min. Benzyl bromide (12.8 g, 74.8mmol) is added, and the solution is stirred for 3 h. The reactionmixture is poured into 0.1 M HCl/ice slurry and extracted with EtOAc(3×150 mL). The combined organic layer is washed with saturated aqueousNaHCO₃ (3×100 mL) and brine (200 mL), dried (MgSO₄), and solvent isevaporated in vacuo. The crude material is purified to afford the benzylester, which is then dissolved in dioxane (20 mL) and stirred at roomtemperature. LiOH.H₂O (1.60 mg, 38 mmol) dissolved in water (10 mL) isadded and the reaction stirred until the benzyl ester had disappeared(by TLC and LCMS). The solvent is removed in vacuo, and the crudematerial is partitioned with EtOAc (50 mL) and 1N HCl (50 mL). Theaqueous layer is extracted with EtOAc (2×50 mL), and the combinedorganic phases are washed with 1M NaHSO₄ (2×50 mL) and brine (50 mL),and dried with MgSO₄. The solvent is evaporated and the crude materialpurified by flash chromatography (EtOAc:Hexanes gradient) to affordReference compound 13 as a white powder.

D-Homophenylalanine ethyl ester hydrochloride (5.00 g, 20.5 mmol) andDIEA (8.7 mL, 51.25 mmol) are dissolved in THF (100 mL), and stirred atroom temperature. Mesyl chloride (1.67 mL, 21.52 mmol) is addeddropwise, and the reaction stirred for 6 hours at room temp. The THF isevaporated and the crude dissolved in EtOAc (100 mL) and washed withwater (100 mL), 1N HCl (2×100 mL) and brine (100 mL), and dried (MgSO₄).The solvent is removed in vacuo, and the crude material purified withflash chromatography (hexanes:EtOAc) to afford the ethyl ester. Theethyl ester is dissolved in dioxane (50 mL), and stirred at roomtemperature. LiOH.H₂O (1.00 mg, 24 mmol) dissolved in water (20 mL) isadded, and the reaction stirred until the ethyl ester had disappeared(by TLC and LCMS). The solvent is removed in vacuo, and the crudematerial is partitioned with EtOAc (50 mL) and 1N HCl (50 mL). Theaqueous layer is extracted with EtOAc (2×50 mL), and the combinedorganic phases are washed with 1M NaHSO₄ (2×50 mL) and brine (50 mL),and dried with MgSO₄. The solvent is evaporated, and the crude materialpurified by flash chromatography (EtOAc:Hexanes gradient) to affordReference compound 14 as a white powder.

Cbz-D-Cys(Bzl)-OH is first prepared from H-D-Cys(Bzl)-OH and Cbz-OSufollowing methods analogous to those described for the preparation ofintermediate compound 1-B. Cbz-D-Cys(Bzl)-OH (296 mg, 0.86 mmol) is thendissolved in 5 mL of MeOH:H₂O (50:50), and oxone (792 mg, 1.29 mmol) isadded. The reaction mixture is stirred overnight at room temperature;diluted with water (10 mL) and extracted with dichloromethane (3×20 mL)land the organic phase is then washed with brine (30 mL) and dried(MgSO₄). The solvent is removed in vacuo to give Reference compound 15,and used without further purification.

Reference compound 16 is prepared starting from D-phenylglycine ethylester hydrochloride following methods analogous to those described forReference compound 14.

In the reaction scheme above for preparing Reference compound 17, thereagents and conditions are: (a) i. t-BuOH, ClSO₂NCO, CH₂Cl₂ 0° C. to rtii. 17-A, CH₂Cl₂, (b) 1 N NaOH in water.

17-B: In a round bottom flask, N-chloroisocyanate (2.5 mL, 28.7 mmol) isdissolved in dichloromethane (150 mL) and cooled to 0° C. Adichloromethane solution of t-BuOH (2.7 mL, 28.7 mmol) is added dropwise. Fifteen minutes following completion of alcohol addition,commercially available 17-A is added in a dichloromethane solution. Thereaction is then allowed to warm to ambient temperature and stir for 3h. The reaction mixture is diluted with water and washed with water; andthe organic layers are separated and dried over MgSO₄ to yield compound17-B as an oil that is used directly in the next step without furtherpurification.

17: The ethyl ester 17-B (1.46 g, 5 mmol) is dissolved in 1N NaOH (15ml, 15 mmol). After the starting material had disappeared (by LCMS), thereaction is acidified with 1 N HCl. The product precipitated as a whitesolid, and used directly in the next step without further purification.

Reference compound 18 is prepared according to the method reported inOrg. Lett. 5:125-128 (2003). Boc-D-homophenylalanine (1.0 g, 3.58 mmol),methylated using Me₂SO₄ and NaH in THF and catalytic water, gave a whitepowder.

Reference compound 19 is prepared from D-homocyclohexylalanine ethylester hydrochloride following methods analogous to those described forReference compound 14.

20-B: 4-piperidine ethanol (20-A) (5.0 g, 39.7 mmol) is dissolved in THF(120 mL). Triethylamine (5.6 mL, 40 mmol) is added, and the solution iscooled to 0° C. Boc₂O (9.59 g, 44 mmol) is added, and the reaction isstirred overnight at room temperature. Solvent is removed in-vacuo; thecrude residue dissolved in ethyl acetate (120 mL) is added; the solutionis washed with 0.1 N HCl (3×100 mL) and brine (1×100 mL); dried withMgSO₄; and filtered and solvent evaporated in vacuo to give compound20-B as a clear oil.

20-C: Trichloroisocyanuric acid (2.66 g, 11.46 mmol) is added to asolution of the alcohol (2.39 g, 10.42 mmol) in DCM, and the solution isstirred and maintained at 0° C., followed by addition of a catalyticamount of TEMPO. After the addition, the mixture is warmed to roomtemperature and stirred for an hour and then filtered on Celite. Theorganic phase is washed with saturated aqueous Na₂CO₃, followed by 1NHCl and brine. The organic layer is dried (MgSO₄), and the solvent isevaporated to yield 20-C. ¹H-NMR (CDCl₃, 400 MHz) δ 9.72 (1H, s),4.07-4.01 (2H, m), 2.70-2.57 (2H, m), 2.35-2.31 (2H, m), 2.05-1.94 (1H,m), 1.64-1.46 (2H, m), 1.39 (9H, s), 1.30-1.02 (2H, m).

20-D: To a solution of Cbz-α-phosphonoglycine trimethyl ester, (2.8 g,8.45 mmol) in THF at −78° C. is added 1,1,3,3-tetramethyl-guanidine(1.022 ml, 8.14 mmol). After 10 minutes, the aldehyde 20-C (1.76 g, 7.76mmol) is added. The solution is then placed in an ice bath at 0° C. for1 hour, and then allowed to warm to room temperature and stirred onemore hour. The solution is diluted with EtOAc, washed with 1M NaHSO₄,dried (MgSO₄) and concentrated in vacuo. The residue is purified bychromatography (ISCO) with Ethyl acetate/Hexane 0 to 100% to afford 20-Das a yellow oil. MS m/z 333.2 (M+1), ¹H NMR (CDCl3, 400 MHz) δ.7.35-7.33 (5H, m), 6.63 (1H, t, J=8 Hz), 6.30 (1H, bs), 5.12 (2H, s),4.10-4.04 (2H, m), 3.73 (3H, s), 2.67-2.62 (2H, m), 2.14 (2H, t, J=6.8Hz), 1.63-1.46 (3H, m), 1.43 (9H, s), 1.14-1.06 (2H, m).

20-E: A Parr vessel is charged with 20-D (1.0 g, 2.31 mmol) and MeOH(100 mL) under nitrogen. The solution is subjected to three cycles ofvacuum and nitrogen bubbling, and the catalyst(R,R)-Ethyl-DuPHOS-Rh(COD) triflate is added (30 mg, 0.04 mmol). Themixture is placed under 60 psi of hydrogen gas at room temperature for24 h. The conversion to 20-E is complete after 24 h with >99% e.e., thesolvent is removed in vacuo, and the crude product is purified by silicagel chromatography (hexanes/EtOAc).

20-F: Intermediate 20-E is dissolved in MeOH, the solution is flushedwith nitrogen, and Pd/Carbon (5% wt, Degussa) is added. The mixture isplaced under 50 psi of hydrogen gas at room temperature and shaken for24 h. The mixture is flushed with nitrogen and filtered through Celite.The cake is washed with MeOH, and the combined organic solution isconcentrated under vacuum. Hexanes is added and then evaporated toazeotrope the remaining methanol to afford 20-F as an oil, which is thenused in the next step without further purification.

20-G: Intermediate 20-F (0.6 g, 1.99 mmol) is dissolved in THF (10 mL),and 2,4,6-collidine (315 mg, 2.38 mmol) and methanesulfonyl chloride(0.170 ml, 2.19 mmol) are added to the solution and stirred for 2 h. Thereaction is diluted with EtOAc (50 mL) solution; washed with 1M NaHSO₄(2×25 mL) and brine (25 mL); and dried (MgSO₄). The solvent is removedin vacuo, and the crude residue purified by flash chromatography using agradient of hexanes and EtOAc to afford the desired product 20-G.

20-H: Compound 20-G (0.70 g, 1.84 mmol) is dissolved in dioxane (7 mL),and LiOH.H₂O (232 mg, 5.55 mmol) dissolved in water (4 mL) is added. Thereaction mixture is stirred for 1 h. The solvent is evaporated; theresidue diluted with EtOAc (25 mL) and washed with 1N NaHSO₄ (25 mL) andbrine (25 mL); and dried (MgSO₄). The solvent is removed in vacuo, andthe crude purified by silica gel chromatography (Hexanes/EtOAc gradient)to afford the desired product, Reference compound 20, as a white solid.

Reference compound 21 is prepared starting from 3-cyanophenylalninefollowing methods analogous to those described for the preparation ofReference compound 14.

Reference compound 22 is prepared from D-homophenylalanine followingmethods analogous to those described for the preparation of Referencecompound 2-B.

Reference compound 23 is prepared from D-homophenylalanine andN-(cyclopropylmethyloxycarbonyloxy)-succinimide following methodsanalogous to those described for the preparation of Reference compound2-B.

Intermediate 24-A is prepared from Reference compound 5-A bydeprotection of the Boc group in 50% TFA in CH₂Cl₂, followed byevaporation of solvent in vacuo. Intermediates 24-B to 24-C are preparedfollowing methods analogous to those described for Reference compound 2.

Reference compound 25 is prepared following methods analogous to thosedescribed for the preparation of Reference compound 24.

Reference compound 26 is prepared from D-homophenylalanine andN-(iso-butyloxycarbonyloxy)-succinimide following methods analogous tothose described for the preparation of Reference compound 2-B.

Reference compound 27 is prepared from D-homophenylalanine andN-(cyclopentyloxycarbonyloxy)-succinimide following methods analogous tothose described for the preparation of Reference compound 2-B.

Reference compound 28 is prepared from D-homophenylalanine andN-(cyclopentlymethyloxycarbonyloxy)-succinimide following methodsanalogous to those described for the preparation of Reference compound2-B.

Reference compound 29 is prepared from D-homophenylalanine andN-(cyclohexyloxycarbonyloxy)-succinimide following methods analogous tothose described for the preparation of Reference compound 2-B.

Reference compound 30 is prepared from D-3-chlorophenylalanine andN-(cyclopentylmethyloxycarbonyloxy)-succinimide following methodsanalogous to those described for the preparation of Reference compound2-B.

Reference compound 31 is prepared from D-3-chlorophenylalanine andN-(cyclohexyloxycarbonyloxy)-succinimide following methods analogous tothose described for the preparation of Reference compound 2-B.

Reference compound 32 is prepared from D-Homophenylalanine ethyl esterhydrochloride and phenoxyacetyl chloride following methods analogous tothose described for the preparation of Reference compound 14.

Reference compound 33 is prepared following methods analogous to thosedescribed for the preparation of Reference compound 24.

Reference compound 34 is prepared from 3-nitrophenylglycine followingmethods analogous to those described for the preparation of Referencecompound 2.

Reference compound 35 is prepared from glycine andN-(cyclohexyloxycarbonyloxy)-succinimide following methods analogous tothose described for the preparation of Reference compound 2-B.

In the above reaction scheme above for preparing Reference compound 36,the reagents and conditions are: (a) PPh₃, DIAD, PhOH, toluene, rt (b)i. TFA, CH₂Cl₂, MeOH rt, 1 h. ii. CyclohexylOCOCl, Pyridine, DMAP, rt,THF.

36-B: To a round bottom flask is charged with commercially available17-A (939 mg, 2.6 mmol), triphenylphosphine (763 mg, 2.8 mmol), phenol(346 mg, 3.8 mmol), toluene (30 mL) and diisopropyldiazodicarboxylate(542 μL, 2.8 mmol) and stirred at ambient temperature. The reactionmixture is concentrated to dryness, and the product is purified from thereaction mixture via silica gel chromatography to afford a white foamused in the preparation of 36-C.

36-C: A 40 mL vial is charged with 36-B (2.0 mmol), dichloromethane, 10mL, methanol (1 mL) and TFA (10 mL). After 1 h at ambient temperature,all volative reagents are removed in vacuo to afford the TFA salt thatis used directly in the next step. The TFA salt andN-(cyclohexylcarbonyloxy)succinimide (2.0 mmol) are added to a roundbottomed flask containing THF (20 mL), pyridine (600 μL) and DMAP (˜10mg, catalytic). The mixture is stirred at room temperature overnight.The clear solution is diluted with EtOAc (200 mL); washed with 1N HCl(3×100 mL) and brine (1×100 mL); and dried with MgSO₄. Solvent isevaporated in vacuo to afford the desired product as a white solid withis used without further purification.

36: The ethyl ester 36-C (2 mmol) is dissolved in 1N NaOH (6 mL, 6mmol). After the starting material had disappeared (by LCMS), thereaction is acidified with 1 N HCl and the product precipitated as awhite solid that is used without further purification.

Intermediate 37-A is prepared from intermediate 2-F (Reference compound2) following similar methods for preparing intermediate 20-B (Referencecompound 20); and hydrogenolysis following analogous methods used forReference compound 1 gives 37-B.

38-B: 2-Chloro-4-nitrotoluene (8.55 g, 50.0 mmol) is dissolved in1,2-dichloroethane (120 mL). N-Bromosuccinimide (9.74 g, 55 mmol) andbenzoyl peroxide (0.25 g, 1.03 mmol) are added, and the reaction mixtureis stirred and heated to reflux for 16 h. The reaction is cooled to roomtemperature, and most of the solvent is evaporated in vacuo, leavingapproximately 30 mL of dichloroethane. Hexanes (30 mL) anddichloromethane (30 mL) are added, and the precipitate that formed isfiltered, and the filtrate washed with hexanes. The combined organicsolution is evaporated in vacuo to afford benzyl bromide 38-B, which isused directly in the next step without further purification.

38-C: Diethyl acetamidomalonate (7.16 g, 33.0 mmol) is dissolved in EtOH(50 mL), and NaOEt (2.25 g, 33.0 mmol) is added. The solution is stirredand heated to 80° C. Bromide 38-B (9.2 g, 37.0 mmol) is dissolved inEtOH (50 mL) and added dropwise to the stirring reaction mixture. After18 h, the solution is cooled to room temperature and a precipitateformed which is filtered. The precipitate is washed with water andrecrystallized with 5% aqueous EtOH, filtered, and dried in vacuo.

38-D: Malonate 38-C is added to 12 N HCl, and the mixture is heated toreflux for 14 h. The reaction mixture is then cooled to 4° C. and aprecipitate formed. The precipitate is washed with 1 N HCl and dried invacuo.

38-F: Intermediate 38-D (4.59 g, 16.33 mmol) and Cbz-OSu (3.99 g, 16.0mmol) are added to a solution of THF (60 mL) and water (20 mL). Et₃N(9.1 mL, 65.32 mmol) is added, and the solution is stirred for 18 h atroom temperature. The solvent is evaporated in vacuo, and the residue istaken up and partitioned between EtOAc (100 mL) and 1N HCl (100 mL). Theorganic phase is washed with 1N HCl (2×100 mL) and brine (100 mL), anddried with MgSO₄. Solvent is removed in vacuo and the crude material isrecrystallized with ethanol and water.

Reference compound 39 is prepared from 5-methyl-2-nitroanisole followingmethods analogous to those described for the preparation of Referencecompound 38.

Reference compound 40 is prepared from 3-methyl-4-nitrobenzyl bromidefollowing methods analogous to those described for Reference compound38.

In the above Scheme 8, the reagents and conditions are: (a)Ethyltrifluoroacetate, K₂CO₃, Dioxane, H₂O, 0° C.; (b) CH₃I, K₂CO₃, DMF,110° C.; (c) LiOH, THF/H₂O, 23° C.; (d) Boc₂O, Et₃N, THF, 23° C.; (e) i.iso-BuOCOCl, Et₃N, THF; ii. NaBH₄, H₂O; (f) Dess-Martin periodinane,CH₂Cl₂; (g) iso-PrMgCl, benzoxazole, THF, −20° C. for 30 min, then −20°C. to rt; (h) H₂, (40 psi), 10% Pd/C, EtOH.

41-A: Cbz-L-Lys-OH (5.6 g, 20.00 mmol) is suspended in water (40 mL) anddioxane (5 mL), and cooled in an ice-bath to 0° C. Potassium carbonate(5.22 g, 40.0 mmol, 2 equiv.) and ethyltrifluoroacetate (7.15 mL, 60.00mmol, 3.0 eq.) are added to the reaction and stirred at 0° C. for 1 hr.After reaction is judged as complete by LC/MS, the reaction is extractedwith diethyl ether (3×100 mL). The aqueous layer is acidified with 100mL 10% citric acid and extracted with ethyl acetate (3×100 mL). Theorganics are combined, washed with saturated NaCl, dried over Na₂SO₄ andevaporated to dryness to provide 41-A as a clear oil; LC/MS [M+H] 377.1,found 377.2.

41-B: This compound is prepared from 41-A using methods described in Xueet al. J. Med. Chem. 2001, 44 (16), 2636.

41-C: This compound is prepared by analogy to Scheme 1 step e.

41-D: Compound 41-C (3.44 g, 11.70 mmol) is dissolved in THF (117 mL,0.1 M) at 23° C. Boc anhydride (5.10 g, 23.40 mmol, 2.0 equiv.) andtriethylamine (3.25 mL, 23.40 mmol, 2.0 equiv.) are added to thereaction. The reaction is monitored to completion by LC/MS. THF isremoved in vacuo, dissolved in ethyl acetate (150 mL) and extracted with1M HCl (2×75 mL) and saturated NaCl; dried over Na₂SO₄; filtered andevaporated to provide 41-D as a clear oil. LC/MS [M+H] 395.2; found395.2.

41-E to 41: These intermediates are prepared following methods analogousto those described for Reference compound 1.

In the above Scheme 9, the reagents and conditions are: (a) Cbz-OSu,Et₃N, THF, H₂O; (b) i. iso-BuOCOCl, Et₃N, THF; ii. NaBH₄, H₂O; (c)Dess-Martin periodinane, CH₂Cl₂; (d) iso-PrMgCl, benzoxazole, THF, −20°C., 30 min, then, −20° C. to rt; (e) Hoveyda-Grubbs metathesis catalyst,4-Methylene-N-Boc-piperidine, DCM, 40° C.; (f) H₂, (40 psi), 10% Pd/C,EtOH.

42-A: This compound is prepared by analogy to Reference compound 38 stepd using L-allylglycine as the amino acid component of the reaction.

42-B-D: These compounds are prepared by analogy to Reference compound 1step a, b and c, respectively.

42-E: Anhydrous dichloromethane (4 mL, 0.2 M) is added via syringe to42-D (270 mg, 0.766 mmol, 1.0 eq.), Hoveyda-Grubbs 2^(nd) Generationmetathesis catalyst(1,3-Bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene)dichloro(o-isopropoxyphenylmethylene) ruthenium II dichloride) (98 mg,0.115 mmol, 15 mol %) under a nitrogen atmosphere.N-Boc-4-methylenepiperidine (0.60 mL, 0.268 mmol, 4.0 eq.) is added viasyringe, and the reaction is fitted with a reflux condenser and heatedto 40° C. for 12 h. After the reaction is judged as complete by LC/MS,the reaction mixture is directly purified by automated silica-gelpurification (0-100% ethyl acetate in hexanes) to provide 42-E as a darkgreen oil. MS m/z 422.3 (M-Boc+1).

42: This compound is prepared by analogy to Reference compound 1 step d.

In the above Scheme 10, the reagents and conditions are: (a)Hoveyda-Grubbs metathesis catalyst, N-Boc-4-amino-1-butene, DCM, 40° C.;(f) H₂, (40 psi), 10% Pd/C, EtOH. Compounds 43A and 43 are prepared byanalogy to Reference compound 42 step e and f, respectively.

Reference compound 44 is prepared from 5-methylbenzoxazole followingmethods analogous to those described for the preparation of Referencecompound 1.

Reference compound 45 is prepared from 5-chlorobenzoxazole followingmethods analogous to those described for the preparation of Referencecompound 1.

Reference compound 46 is prepared from 5-fluorobenzoxazole followingmethods analogous to those described for the preparation of Referencecompound 1.

Reference compound 47 is prepared from 6-fluorobenzoxazole followingmethods analogous to those described for the preparation of Referencecompound 1.

In the above Scheme 11, the reagents and conditions are: (a) TFA/CH₂Cl₂75:25, 23° C.; (b)N,N-Bis(tert-butoxycarbonyl)-1H-pyrazole-1-carboxamidine, DIEA, MeOH;(c) H₂, (40 psi), 10% Pd/C, EtOH. Intermediate 48-A is prepared inanalogy to Scheme 1, step a using Compound 42-A. Intermediate 48-B isprepared in analogy to Reference compound 2 step f. Intermediate 48 isprepared in analogy to Reference compound 1 step d.

Reference compound 49 is prepared from 6-methylbenzoxazole followingmethods analogous to those described for the preparation of Referencecompound 1.

Reference compound 50 is prepared fromN-(cyclohexyloxycarbonyloxy)-succinimide following methods analogous tothose for Reference compound 2-B.

Reagents and conditions are as follows: (a) Cbz-OSu, Et₃N, THF, water;(b) Hoveyda-Grubbs metathesis catalyst, N-Boc-4-methylenepiperidine,DCM, 40° C. Compound 51-A is prepared by analogy to Scheme 2 step a forpreparing Reference compound 2. Compound 51 is prepared by analogy toScheme 9 step e for preparing Reference compound 42.

In the above reaction scheme for preparing Reference compound 52, thereagents and conditions are: (a) Cbz-OSu, Et₃N, THF, Water; (b)Hoveyda-Grubbs metathesis catalyst, N-Boc-4-methylenepiperidine, DCM,40° C. Intermediate 52-A is prepared following methods analogous tothose described for Reference compound 2 step a. Reference compound 52is prepared following methods analogous to those described for Referencecompound 42 step e.

1-Boc-homopiperazine (1.00 g, 4.2 mmol) and DIEA (8.7 mL, 51.25 mmol)are dissolved in THF (15 mL) and stirred at room temperature. Mesylchloride (1.67 mL, 21.52 mmol) is added dropwise, and the reactionstirred 6 h at room temperature. The reaction is diluted with EtOAc (100mL); washed with 1N HCl (2×50 mL), saturated aqueous NaHCO₃ (50 mL) andbrine (50 mL); dried with MgSO₄; and then the solvent is evaporated todryness in vacuo. The crude residue is taken up in 50% TFA in DCM andstirred for 1 h, and then evaporated to dryness in vacuo to affordReference compound 53 which is used without further purification.

(2S,4R)-1-((benzyloxy)carbonyl)-4-(piperidine-1-carboxyloyloxy)pyrrolidine-2-carboxylicacid is prepared starting from L-Hydroxyproline methyl ester followingmethods analogous to those described for the preparation of Compound1-G.

Reference compound 55 is prepared starting from D-Allylglycine followingmethods analogous to those described for the preparation of Referencecompound 2-B.

Reference compound 56 is prepared starting from D-Allylglycine followingmethods analogous to those described for the preparation of Referencecompound 2-B.

57-B: A solution of L-4-pyridylalanine 57-A (1.0 g), 10% Pd/C (300 mg)in EtOH (40 mL) and 1 N HCl aqueous solution (20 mL) is shaken on ParrShaker for 12 hours under a hydrogen atmosphere at 50 psi. The catalystis filtered and the filtrate is concentrated to afford the piperidine57-B.

57-C: Piperidine 57-B (1.0 g) and copper sulfate pentahydrate (1.5 g)are dissolved in dioxane (50 mL) and water (30 mL). The pH of thesolution is adjusted to pH 9 with 2 N sodium hydroxide at roomtemperature. Di-t-butyl-dicarbonate (2.63 g) is then added. The slurrymixture is stirred for overnight at room temperature. The precipitationis collected and washed with water, and then dissolved in dioxane. ThepH of the solution is adjusted with 4N sodium hydroxide to pH 12 andCbz-OSu (3.2 g) is added. The mixture is stirred overnight. Dioxane isremoved under reduced pressure. The residue is acidified with 1Nhydrochloric acid to pH 2-3, and extracted with ethyl acetate. Thecombined ethyl acetate solution is washed with brine, dried over sodiumsulfate, and concentrated to dryness to give an oil. This crude materialis taken up in DCM (50 mL) and MeOH (10 mL), and a 2 M solution ofTMSCHN₂ in ether (3.5 mL) is added until solution is light yellow. Thesolvents are then removed under reduced pressure. The residue ispurified on silica gel chromatography with 20% ethyl acetate in hexaneto give the α-N-Cbz, ε-N-Boc methyl ester 57C.

57-D: A solution of 57-C (150 mg) in dichloromethane (20 mL) is cooledto −78° C., then 1M DiBAl-H in hexane (1.07 mL) is added over a periodof 10 min. The mixture is stirred at −78° C. for 50 min, and a 5% citricacid aqueous solution (10 mL) is added to quench the reaction. The DCMlayer is separated, and the aqueous layer is extracted with DCM. Thecombined DCM solution is washed with water, dried over sodium sulfateand filtered. The filtrate is concentrated to dryness to give thealdehyde. The aldehyde is dissolved in THF (8 mL) and used directly inthe next step.

57-E: To a solution of benzoxazole (128 mg) in THF, 2.5 M BuLi in hexaneis added at −30° C. The solution is stirred at −20° C. for 40 min, and adark red solution is obtained. The aldehyde 57-E in THF (8 mL) is thenadded to the dark red solution at −30° C. over a period of 5 min. Themixture is stirred for 2 h at −10° C. Saturated aqueous ammoniumchloride solution (10 mL) is added to the reaction mixture to quench thereaction. THF is removed under reduced pressure. The residue isextracted with ethyl acetate. The combined ethyl acetate solution iswashed with 1N HCl aqueous solution, water, saturated aqueous sodiumbicarbonate, and brine respectively, and then dried over anhydroussodium sulfate and filtered. The filtrate is concentrated, and theresidue is purified by silica gel chromatography with 30% to 50% ethylacetate in hexane to give the desired product.

57-F: Intermediate 57-E is dissolved in MeOH and Pd/C (50 mg) is added.Hydrogen gas from a balloon is bubbled through the mixture for 30 min.The catalyst is then removed by filtration, and the filtrate isconcentrated to give the amine.

EXAMPLE 1

1-B: Finely powdered KOH (19.4 g, 0.346 mol) is dissolved in DMSO andstirred at room temperature for 20 min. and then cooled to 0° C.N-Boc-trans-4-hydroxy-L-proline (Boc-Hyp-OH, 1-A) (10 g, 43.3 mmol) isdissolved in DMSO (10 mL) and added, and the reaction mixture is stirredfor an additional 10 min at 0° C. Next, 4-chlorobenzyl chloride (33 g,0.204 mol) is added, and the reaction mixture is stirred at 0° C. for anadditional 15 min, after which point the ice bath is removed, and thereaction mixture is allowed to warm to room temperature and stir for 4h. The reaction mixture is poured into water (300 mL), and the reactionvessel is rinsed with an additional aliquot of water (300 mL). Thecombined aqueous layer is extracted with ether (2×300 mL) and discarded.The aqueous layer is acidified with 87% H₃PO₄ to pH 2-3, and thenextracted with ether (3×300 mL). The combined ether extracts are washedwith water (2×400 mL) and brine (2×400 mL); dried over MgSO₄; filteredand concentrated in vacuo. The residue is purified by chromatography onsilica gel with EtOAc/Hexanes (gradient 0 to 100%) to yield the compound1-B as a clear oil. MS m/z 256.1 (M+1-Boc); ¹H-NMR (DMSO-D₆, 400 MHz) δ7.39-7.31 (4H, m), 4.52-4.40 (2H, m), 4.16-4.10 (2H, m), 3.48-3.41 (2H,m), 2.40-2.30 (1H, m), 2.03-1.94 (1H, m), 1.39-1.34 (9H, m).

1-C: A solution of (trimethylsilyl)diazomethane (2M in diethylether)(4.7 ml, 9.45 mmol) is added to carboxylic acid 1-B (2.4 g, 8.6 mmol)dissolved in DCM/MeOH 5:1 (25 mL). When the starting material isconsumed as determined by LCMS, the reaction mixture is concentrated invacuo, and the crude residue is purified by flash chromatography(gradient EtOAc:Hexanes) to afford methyl ester as a clear oil.

1-D: A round bottomed flask is charged with a stirbar and 1-C (510 mg,1.38 mmol). TFA (50%) in DCM (6 mL) is added, and the solution isstirred for 1 h at room temperature. The solvent is removed in vacuo,hexanes are added and then evaporated again in vacuo to dryness, andrepeated if necessary to azeotrope remaining TFA. The crude material isused directly in the next step without further purification.

1-E: The crude material is dissolved in DCM (10 mL) followed by additionof Cbz-D-homoPhe-OH (Reference compound 22) (432 mg, 1.38 mmol) and HATU(577 mg, 1.52 mg), and the solution is stirred at room temperature for10 min. DIEA (0.72 mL, 4.14 mmol) is added to the solution, and thereaction mixture is allowed to stir overnight at room temperature. Thesolvent is removed in vacuo, and the crude material is directly purifiedby flash chromatography (40 g silica, hexanes/EtOAc gradient). Thesolvent is removed in vacuo to afford the desired compound as an oilysemisolid.

1-F: Methyl ester 1-E (756 mg, 1.34 mmol) is dissolved in dioxane (10mL). Lithium hydroxide monohydrate (84 mg, 2.00 mmol) is dissolved inwater (5 mL) and added dropwise to the solution of methyl ester 1-E, andallowed to stir overnight. The reaction mixture is concentrated in vacuoto remove dioxane and then acidified with 1M NaHSO₄. This is extractedwith EtOAc, and the combined organic layer is washed with brine anddried with MgSO₄. The solvent is removed in-vacuo to afford carboxylicacid 1-F as a waxy solid.

1-G: Carboxylic acid 1-F (534 mg, 0.97 mmol) is dissolved in DCM (18mL). Reference compound 1 (3.38 mg, 0.97 mmol) and HATU (405 mg, 1.07mmol) are added, and the mixture is stirred for 10 min at roomtemperature. Next, DIEA (0.51 mL, 2.91 mmol) is added, and the reactionmixture is left to stir overnight at room temperature. The solvent isremoved in vacuo, the crude is redissolved in EtOAc (50 mL) and washedwith 1M HCl (2×25 mL), followed by saturated aqueous NaHCO₃ (2×25 mL)and brine (25 mL), and dried with anhydrous Na₂SO₄. Solvent is removedto afford the 1-G as a white foam, which is purified by flashchromatography (Hexanes/EtOAc) to afford the desired product.

1-H. Alcohol 1-G (727 mg, 0.82 mmol) is dissolved in DCM (10 mL) andDess-Martin periodinane (524 mg, 1.24 mmol) is added. The reactionmixture is stirred overnight at room temperature. The solvent is removedin vacuo, and the crude is purified by flash chromatography (40 g silicacolumn) using a gradient of EtOAc:Hexanes to afford the ketone 1-H as awhite foam.

1-I: 1-H (579 mg, 0.66 mmol) is dissolved in DCM (1 mL), and TFA 50% inDCM (5 mL) is added. The reaction is stirred at room temp for 2 h, andthe solvent is removed in vacuo. The crude material is purified byreverse-phase HPLC, and the solvent is lyophilized to afford as theproduct as a white powder.

EXAMPLES 2-46

Examples 2-46 are obtained by repeating the procedures described inExample 1, using appropriate Reference compounds and reagents whichwould be apparent to those skilled in the art, for example:

Example 2, using Reference compound 3;

Example 3, using Reference compound 14;

Example 4, using Reference compound 3 and Reference compound 14;

Example 5, using Reference compound 16;

Example 6, using Reference compound 21;

Example 7, using N-p-tosylglycine;

Example 8, using N-mesyl-L-alanine;

Example 9, using N-mesylglycine;

Example 10, using Reference compound 19;

Example 11, using D-homocyclohexylalanine (D-homoCha);

Example 12, using trans-3-hydroxy-L-proline;

Example 13, using trans-3-hydroxy-L-proline and Reference compound 4;

Example 14, using Reference compound 4;

Example 15, using Reference compound 4;

Example 16, using trans-3-hydroxy-L-proline and Reference compound 4;

Example 17, using Reference compound 23;

Example 18, using trans-3-hydroxy-L-proline and Reference compound 23;

Example 25, using Reference compound 14;

Example 26, using trans-3-hydroxy-L-proline;

Example 27, using trans-3-hydroxy-L-proline and Cbz-D-cyclohexylalanine;

Example 28, using trans-3-hydroxy-L-proline andCbz-D-3-trifluoromethylphenylalanine;

Example 30, using trans-3-hydroxy-L-proline and Reference compound 14;

Example 31, using Reference compound 14;

Example 32, using Reference compound 12;

Example 34, using O-benzyl-D-tyrosine;

Example 35, using O-benzyl-D-serine;

Example 36, using D-homocyclohexylalanine (D-homoCha);

Example 38, using D-phenylglycine;

Example 39, using Reference compound 2;

Example 40, using Reference compound 8;

Example 41, using Reference compound 9;

Example 42, using Reference compound 2;

Example 43, using Reference compound 24;

Example 44, using Reference compound 2;

Example 45, using Reference compound 5; and

Example 46, using Reference compound 25.

EXAMPLE 47

47-B: This compound is prepared from Cbz-Hyp-OH and cyclohexylbromidefollowing methods analogous to those described for the preparation ofintermediate 1-B in Example 1.

47-C: This compound is prepared from 47-B and Reference compound 1following methods analogous to those described for the preparation ofintermediate 1-G in Example 1.

47-D: A Parr reaction vessel is charged with 47-C (2.5 g, 3.6 mmol),Pd/C (3.6 g, 0.36 mmol, 1 eq), t-BuOH (20 mL) and water (5 mL). Thevessel is placed into a Parr apparatus and shaken for 18 h under apressure of 50 psi of H₂ gas. The reaction mixture is filtered through apad of Celite, and the volatile solvents are removed under reducedpressure to afford compound 47-D which is used directly in the next stepwithout further purification.

47-E: A 40 mL vial is charged with 47-D (75 mg, 0.13 mmol), N-Mesyl(D)-homophenylalanine (56 mg, 0.15 mmol, 1.1 eq), HATU (75 mg, 0.13mmol, 1.1 eq), iPr₂NEt (0.03 ml, 0.16 mmol, 1.2 eq) and CH₂Cl₂ (2 mL).The reaction is stirred at room temperature for 1 h. The volatilereagents are removed under reduced pressure, and the reaction mixture isdissolved in EtOAc. The organic layers are washed with NaHSO₄, watersaturated NaHCO₃, and brine. The organic layers are then dried withMgSO₄, and the product purified from the reaction mixture via silica gelchromatography using a gradient of 3-9% MeOH in CH₂Cl₂ over gradient toafford compound 47-D.

47-F: This compound is prepared by oxidation of 47-E following methodsanalogous to those described for the preparation of intermediate 1-H inExample 1.

47-G: This compound is prepared by deprotection of 47-F followingmethods analogous to those described for the preparation Example 1.

EXAMPLES 48-115

Examples 48-57, 83-84 and 86-87 are obtained by repeating the proceduresdescribed in Example 47, using appropriate Reference compounds andreagents which would be apparent to those skilled in the art, forexample:

Example 48, using D-cyclohexylglycine;

Example 49, using 4-(trifluoromethoxy)-DL-phenylalanine;

Example 50, using Reference compound 23;

Example 51, using Reference compound 27;

Example 52, using Reference compound 28;

Example 53, using D-3-chlorophenylalanine;

Example 54, using 3-(trifluoromethyl)-D-phenylalanine;

Example 55, using Reference compound 29;

Example 56, using Reference compound 30;

Example 57, using Reference compound 31;

Example 83, using Reference compound 36;

Example 84, using Reference compound 17;

Example 86, using Reference compound 26; and

Example 87, using Reference compound 22.

Examples 58-82, 85 and 88-115 are obtained by repeating the proceduresdescribed in Example 1, using appropriate Reference compounds andreagents which would be apparent to those skilled in the art, forexample:

Example 58, using Reference compound 32;

Example 59, using Reference compound 2 and Reference compound 28;

Example 60, using Reference compound 11 and Reference compound 28;

Example 61, using Reference compound 33 and Reference compound 28;

Example 62, using Reference compound 29;

Example 63, using D-pyroglutamic acid;

Example 64, using Reference compound 18;

Example 65, using Reference compound 18 and Reference compound 2;

Example 66, using Reference compound 18 and Reference compound 3;

Example 67, using Reference compound 18 and Reference compound 5;

Example 68, using Reference compound 18 and Reference compound 9;

Example 69, using Reference compound 14 and Reference compound 2;

Example 70, using Reference compound 18 and Reference compound 7;

Example 71, using Reference compound 18 and Reference compound 4;

Example 72, using Reference compound 14 and Reference compound 4;

Example 73, using Reference compound 18 and Reference compound 11;

Example 74, using Reference compound 14 and Reference compound 5;

Example 75, using Reference compound 14 and Reference compound 9;

Example 76, using Reference compound 18 and Reference compound 8;

Example 77, using Reference compound 18 and Reference compound 34;

Example 78, using Reference compound 18 and Reference compound 6;

Example 79, using Reference compound 18 and Reference compound 10;

Example 80, using Reference compound 27;

Example 81, using Reference compound 23;

Example 82, using Reference compound 35;

Example 85, using Boc-D-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid(Boc-D-Tic-OH);

Example 88, using Reference compound 20;

Example 89, using Reference compound 20 and Reference compound 3;

Example 90, using Reference compound 20 and Reference compound 2;

Example 91, using Reference compound 20 and Reference compound 5;

Example 92, using Reference compound 20 and Reference compound 9;

Example 93, using Reference compound 20 and Reference compound 6;

Example 94, using Reference compound 20 and Reference compound 10;

Example 95, using Reference compound 20 and Reference compound 11;

Example 96, using Reference compound 20 and Reference compound 7;

Example 97, using Reference compound 2 and Reference compound 18;

Example 98, using Reference compound 37;

Example 99, using Reference compound 38;

Example 100, using Reference compound 39 and Reference compound 14;

Example 101, using Reference compound 40 and Reference compound 18;

Example 102, using Reference compound 38 and Reference compound 18;

Example 103, using Reference compound 34;

Example 104, using Reference compound 40;

Example 105, using Reference compound 38;

Example 106, using Reference compound 41;

Example 107, using Reference compound 42;

Example 108, using Reference compound 43;

Example 109, using Reference compound 44;

Example 110, using Reference compound 45;

Example 111, using Reference compound 46;

Example 112, using Reference compound 47;

Example 113, using Reference compound 48;

Example 114, using Reference compound 48, with the final compoundisolated as a mixture of diastereomers; and

Example 115, using Reference compound 49 and Reference compound 18.

EXAMPLE 116

In Example 116, the reagents and conditions are: (a) Ag₂O, allylbromide,Et₃N, acetone, 23° C.; (b) TFA, DCM, 23° C. (c) HATU, DIEA, DCM,Reference compound 50, 23° C.; (d) LiOH, dioxanes, water, 23° C.; (e)HATU, DIEA, DCM, Reference compound 1, 23° C.; (f) Dess-Martinperiodinane, DCM; (g) TFA, DCM, 23° C.; (h) Hoveyda-Grubbs metathesiscatalyst, methylenecyclopentane, DCM, 40° C.

Compound 116-A is prepared following procedures in Park, M.-S. J. Kor.Chem. Soc. 45:549 (2001). Compound 116-B is prepared following Example 1step h. Compound 116-C is prepared following Example 1 step d usingReference compound 23 as the acid component. Compound 116-D is preparedfollowing Example 1 step 3. Compound 116-E is prepared following Example1 step f using Reference compound 1 as the amine component. Compound116-F and 116-G are prepared following Example 1 step g and step h,respectively. The compound 116 is prepared following Scheme 9 step e forpreparing Reference compound 42.

EXAMPLE 117

In Example 117, the reagents and conditions are: (a) Hoveyda-Grubbsmetathesis catalyst, methylenecyclopentane, DCM, 40° C.; (b) H₂ (40psi), MeOH, Pd/C (10 wt %, wet); (c) Dess-Martin periodinane, DCM; (d)TFA, DCM, 23° C.

Compound 117-A is prepared following Example 1 step f using Referencecompound 1 as the amine component. Compound 117-B and 117-C are preparedfollowing Example 1 step g and step h, respectively. Compound 117 isprepared following Scheme 9 step e for preparing Reference compound 42.

EXAMPLES 118-123

Examples 118-123 are obtained by repeating the above procedures, usingappropriate Reference compounds and reagents which would be apparent tothose skilled in the art, for example:

Example 118, using styrene in Example 117 step a;

Example 119, using 4-chlorostyrene in Example 116 step h;

Example 120, using N-Boc-L-3-hydroxyproline in Example 116 step a, andmethylenecyclohexane in Example 116 step h;

Example 121, using N-Boc-L-3-hydroxyproline in Example 116 step a, andmethylenecyclohexane in Example 117 step a;

Example 122, using N-Boc-L-3-hydroxyproline in Example 116 step a, andN-Boc-4-methylenepiperidine in Example 117 step a;

Example 123, following methods analogous to those described in Example1, using Reference compound 50 in step d; and

Example 123, following methods analogous to those described for Example1, using Reference Compound 50 in step d.

EXAMPLE 124

In Example 124, the reagents and conditions are: (a) HATU, DIEA, DCM,Reference compound 21, 23° C.; (b) LiOH, dioxanes, water, 23° C.; (c)HATU, DIEA, DCM, Reference compound 1, 23° C.; (d) Dess-Martinperiodinane, DCM; (e) Hoveyda-Grubbs metathesis catalyst,methylenecyclopentane, DCM, 40° C.; (f) TFA, DCM, 23° C.

Compound 124-A is prepared following Example 1 step d using Referencecompound 21 as the acid component and compound 1-A as the aminecomponent. Compound 124-B is prepared following Example 1 step e.Compound 124-C is prepared following Example 1 step f using Referencecompound 1 as the amine component. Compound 124-D is prepared followingExample 1 step g. Compound 124-E is prepared following Scheme 9 step efor preparing Reference compound 42. Compound 124 is prepared followingExample 1 step h.

EXAMPLES 125-134

Examples 125, 127-130 and 133-134 are obtained by repeating theprocedures described in Example 124, using appropriate Referencecompounds and reagents which would be apparent to those skilled in theart, for example:

Example 125, using methylenecyclohexane as a reagent;

Example 127, using 4-carboxystyrene as a reagent;

Example 128, using 4-chlorostyrene as a reagent;

Example 129, using N-Boc-4-methylenepiperidine as a reagent;

Example 130, using methylenecyclopentane as a reagent; and

Example 133, using methylenecyclohexane as a reagent.

Examples 126 and 131-132 are obtained by repeating the proceduresdescribed in Example 1, using appropriate Reference compounds andreagents which would be apparent to those skilled in the art, forexample:

Example 126, using Reference Compound 51;

Example 131, using Reference Compound 52; and

Example 132, using Reference Compound 52.

EXAMPLE 135

135-A: This compound is synthesized from 47-D andCbz-D-homophenylalanine, following methods analogous to those describedfor the preparation of 47-E.

135-B: The Cbz protecting group is removed by hydrogenolysis, usingconditions analogous to those described for the preparation of Referencecompound 1-E.

135-C: Amine 135-B (79 mg, 0.11 mmol) and pyridine (0.2 mL) aredissolved in CH₂Cl₂ (10 mL). 4-fluorophenyl chloroformate (21 mg, 0.12mmol) is added, and the solution is stirred at room temperature forseveral hours until the starting material is consumed (by LCMS). Thesolvent is evaporated and the residue taken up in EtOAc (30 mL). Theorganic phase is washed with 1M NaHSO₄ (2×25 mL) and brine (25 mL), andthen dried with MgSO₄. The solvent is evaporated, and the crude materialpurified by silica gel chromatography (Hexanes/EtOAc gradient 0 to100%).

135-D: This compound is prepared from 135-C, following methods analogousto those described for the preparation of 47-F.

135-E: This compound is prepared from 135-D, following methods analogousto those described for the preparation of 47-G.

EXAMPLES 136-152 AND 258

Examples 136 and 137 are prepared, following methods analogous to thosedescribed in Example 135 and Example 1, respectively. By repeating theprocedures described in the above examples and, using appropriatestarting materials, Examples 138-152 and 258 are obtained.

Table 1 shows compounds of Formula (1), as described in Examples 1-152and 258.

TABLE 1 Physical Data MS (m/z), Elemental Analysis, and Structure ¹H NMR400 M Hz (DMSO-d₆)  1

MS m/z 780.4 (M + 1); Anal. Calcd. for C₄₅H₄₇ClF₃N₅O₉ (1 TFA): C, 60.43;H, 5.30; N, 7.83; Found: C, 59.92; H, 5.09; N, 7.62; ¹H NMR (CD₃CN, 400M Hz) δ 7.91(1H, d, J=8.0 Hz), 7.75 (1H, dd, J=8.0, 3.2 Hz),7.63-7.60(1H, m), 7.54- 7.50(1H, m), 7.38(2H, d, J=4.8 Hz), 7.35-7.24(10H, m), 7.20-7.15(2H, m), 5.54-5.30(1H, m), 5.16-5.01(2H, m),4.49-4.13(5H, m), 3.88-3.73(1H, m), 3.58-3.49(1H, m), 2.92-2.91(1H, m),2.79-2.74 (1H, m), 2.63-2.58(1H, m), 2.26-2.19(1H, m), 2.09- 1.79(1H,m), 1.91-1.84(2H, m), 1.78-1.64(3H, m), 1.61-1.35(4H, m).  2

MS m/z 824.1 (M + 1); Anal. Calcd. for C₄₃H₅₁Cl₂N₇O₈S (1 HCl, 2 H₂O): C,57.58; H, 5.73; N, 10.93; Found: C, 57.11; H, 5.75; N, 10.27; ¹H NMR(CD₃CN, 600 M Hz) δ 8.17(1H, d, J=7.8 Hz), 8.06(1H, d, J=7.8 Hz),8.03-8.01(1H, m), 7.66-7.55(2H, m), 7.41-7.21(10H, m), 7.20-7.12 (3H,m), 5.56-5.50(1H, m), 5.11(1H, d, J=12.6 Hz), 5.03(1H, d, J=12.6 Hz),4.59(1H, t, J=7.8 Hz), 4.44-4.26(3H, m), 3.77-3.71(2H, m), 3.70-3.67(2H, m), 3.57-3.53(2H, m), 3.25-3.10(2H, m), 2.78- 2.58(2H, m),2.32-2.24(1H, m), 1.93-1.86(2H, m), 1.85-1.77(1H, m), 1.76-1.68(1H, m). 3

MS m/z 724.3 (M + 1)  4

MS m/z 768.2 (M + 1)  5

MS m/z 696.8 (M + 1)  6

MS m/z 660.3 (M + 1); Anal. Calcd. for C₃₃H₄₃ClF₃N₅O₁₁S (2 TFA, 1 H₂O):C, 48.92; H, 5.35; N, 8.64; Found: C, 49.22; H, 5.06; N, 8.48; ¹H NMR(CD₃CN, 400 M Hz) δ 7.91(1H, d, J=8.0 Hz), 7.76(1H, d, J=8.4 Hz),7.65-7.39(3H, m), 7.38-7.26(3H, m), 5.87-5.73(1H, m), 5.52-5.42(1H, m),5.22-5.02(2H, m), 4.56-4.36(3H, m), 4.34-4.22 (2H, m), 3.88-3.68(2H, m),2.92(3H, s), 2.44-2.22 (6H, m), 2.17-1.98(2H, m), 1.81-1.54(6H, m).  7

MS m/z 696.2 (M + 1); Anal. Calcd. for C₃₆H₄₅ClF₃N₅O₁₂S (1 TFA, 3 H₂O):C, 50.03; H, 5.25; N, 8.10; Found: C, 50.42; H, 4.71; N, 7.70; ¹H NMR(CD₃CN, 400 M Hz) δ 7.93(1H, d, J=8.0 Hz), 7.78(1H, d, J=8.0 Hz),7.76(2H, d, J=8.0 Hz), 7.66(2H, app t), 7.60-7.51(1H, m), 7.39-7.31 (4H,m), 7.15-7.08(1H, m), 5.56-5.48(1H, m), 4.54- 4.38(3H, m), 4.31-4.24(1H,m), 3.79(2H, d, J=4.0 Hz), 3.66(1H, d, J=11.0, 4.4 Hz), 3.58(1H, d, J=11.2 Hz), 3.02(2H, br s), 2.44(3H, s), 2.33-2.24 (2H, m), 2.21-2.09(2H,m), 1.84-1.59(4H, m).  8

MS m/z 634.2 (M + 1)  9

MS m/z 620.5 (M + 1)  10

MS m/z 730.4 (M + 1)  11

MS m/z 786.4 (M + 1)  12

MS m/z 824.3 (M + 1)  13

MS m/z 774.2 (M + 1)  14

MS m/z 808.4 (M + 1); ¹H NMR (CD₃CN, 400 M Hz) δ 7.85-7.79(1H, m),7.58(1H, d, J=8.4 Hz), 7.50-7.46(1H, m), 7.42-7.38(1H, m), 7.30-7.04(14H, m), 6.10-5.94(1H, m), 5.53-5.42(1H, m), 5.12-4.92(2H, m),4.57-4.42(1H, m), 4.38-4.18(2H, m), 4.17-4.05(1H, m), 3.69-3.54(1H, m),3.37-3.15 (2H, m), 3.14-2.98(2H, m), 2.77-2.48(2H, m), 2.28- 2.14(1H,m), 2.13-2.00(2H, m), 1.78-1.58(4H, m).  15

MS m/z 842.3 (M + 1); ¹H NMR (CD₃CN, 400 M Hz) δ 7.85-7.79(1H, m),7.58(1H, d, J=8.4 Hz), 7.50-7.46(1H, m), 7.42-7.38(1H, m), 7.30-7.04(14H, m), 6.10-5.94(1H, m), 5.53-5.42(1H, m), 5.12-4.92(2H, m),4.57-4.42(1H, m), 4.38-4.18(2H, m), 4.17-4.05(1H, m), 3.69-3.54(1H, m),3.37-3.15 (2H, m), 3.14-2.98(2H, m), 2.77-2.48(2H, m), 2.28- 2.14(1H,m), 2.13-2.00(2H, m), 1.78-1.58(4H, m).  16

MS m/z 698.3 (M + 1)  17

MS m/z 660.3 (M + 1)  18

MS m/z 660.3 (M + 1)  19

MS m/z 814.3 (M + 1); ¹H NMR (CD₃CN, 400 M Hz) δ 7.91(1H, d, J=8.0 Hz),7.75(1H, dd, J= 8.0, 3.2 Hz), 7.63-7.60(1H, m), 7.54-7.50(1H, m),7.38(2H, d, J=4.8 Hz), 7.35-7.24(10H, m), 7.20- 7.15(2H, m),5.54-5.30(1H, m), 5.16-5.01(2H, m), 4.49-4.13(5H, m), 3.88-3.73(1H, m),3.58-3.49(1H, m), 2.92-2.91(1H, m), 2.79-2.74(1H, m), 2.63-2.58 (1H, m),2.26-2.19(1H, m), 2.09-1.79(1H, m), 1.91- 1.84(2H, m), 1.78-1.64(3H, m),1.61-1.35(4H, m).  20

MS m/z 760.4 (M + 1); ¹H NMR (CD₃CN, 400 M Hz) δ 7.91(1H, d, J=8.0 Hz),7.75(1H, dd, J= 8.0, 3.2 Hz), 7.63-7.60(1H, m), 7.54-7.50(1H, m),7.38(2H, d, J=4.8 Hz), 7.35-7.24(10H, m), 7.20- 7.15(2H, m),5.54-5.30(1H, m), 5.16-5.01(2H, m), 4.49-4.13(5H, m), 3.88-3.73(1H, m),3.58-3.49(1H, m), 2.92-2.91(1H, m), 2.79-2.74(1H, m), 2.63-2.58 (1H, m),2.26(3H, s), 2.25-2.19(1H, m), 2.09-1.79 (1H, m), 1.91-1.84(2H, m),1.78-1.64(3H, m), 1.61- 1.35(4H, m).  21

MS m/z 764.4 (M + 1); ¹H NMR (CD₃CN, 400 M Hz) δ 7.91(1H, d, J=8.0 Hz),7.75(1H, dd, J= 8.0, 3.2 Hz), 7.63-7.60(1H, m), 7.54-7.50(1H, m),7.38(2H, d, J=4.8 Hz), 7.35-7.24(10H, m), 7.20- 7.15(2H, m),5.54-5.30(1H, m), 5.16-5.01(2H, m), 4.49-4.13(5H, m), 3.88-3.73(1H, m),3.58-3.49(1H, m), 2.92-2.91(1H, m), 2.79-2.74(1H, m), 2.63-2.58 (1H, m),2.26-2.19(1H, m), 2.09-1.79(1H, m), 1.91- 1.84(2H, m), 1.78-1.64(3H, m),1.61-1.35(4H, m).  22

MS m/z 814.3 (M + 1); ¹H NMR (CD₃CN, 400 M Hz) δ 7.91(1H, d, J=8.0 Hz),7.75(1H, dd, J= 8.0, 3.2 Hz), 7.63-7.60(1H, m), 7.54-7.50(1H, m),7.38(2H, d, J=4.8 Hz), 7.35-7.24(10H, m), 7.20- 7.15(2H, m),5.54-5.30(1H, m), 5.16-5.01(2H, m), 4.49-4.13(5H, m), 3.88-3.73(1H, m),3.58-3.49(1H, m), 2.92-2.91(1H, m), 2.79-2.74(1H, m), 2.63-2.58 (1H, m),2.26-2.19(1H, m), 2.09-1.79(1H, m), 1.91- 1.84(2H, m), 1.78-1.64(3H, m),1.61-1.35(4H, m).  23

MS 830.6 m/z (M + 1); Anal. Calcd. for C₄₃H₄₅BrF₃N₅O₉S (1 TFA): C,54.66; H, 4.80; N, 7.41; Found: C, 54.78; H, 4.57; N, 7.28; ¹H NMR(CD₃CN, 600 M Hz) δ 8.05(1H, br s), 7.94(1H, d, J= 7.8 Hz), 7.78(1H, d,J=8.4 Hz), 7.65-7.61(2H, m), 7.54(1H, t, J=7.2 Hz), 7.45-6.94(9H, m),6.89 (1H, d, J=7.2 Hz), 5.32-5.30(1H, m), 5.15(1H, d, J= 12.6 Hz),5.10(1H, d, J=12.6 Hz), 4.49-4.33(3H, m), 4.22(1H, br s), 3.77(1H, dd,J=11.4, 4.8 Hz), 3.53(1H, d, J=11.4 Hz), 2.97(1H, br s), 2.81-2.73 (2H,m), 2.69-2.57(5H, m), 2.29-2.20(1H, m), 2.13- 1.99(2H, m), 1.95-1.89(2H,m), 1.82-1.64(2H, m), 1.63-1.46(2H, m).  24

MS m/z 830.2 (M + 1); ¹H NMR (CD₃CN, 600 M Hz) δ 7.94(1H, d, J=7.8 Hz),7.78(1H, d, J=8.4 Hz), 7.65-7.61(2H, m), 7.54(1H, t, J=7.2 Hz),7.53-6.79(9H, m), 6.89(1H, d, J=7.2 Hz), 5.32- 5.30(1H, m), 5.15(1H, d,J=12.6 Hz), 5.10(1H, d, J=12.6 Hz), 4.49-4.33(3H, m), 4.22(1H, br s),3.77 (1H, dd, J=11.4, 4.8 Hz), 3.53(1H, d, J=11.4 Hz), 2.97(1H, br s),2.81-2.73(2H, m), 2.69-2.57(5H, m), 2.29-2.20(1H, m), 2.13-1.99(2H, m),1.95-1.89 (2H, m), 1.82-1.64(2H, m), 1.63-1.46(2H, m).  25

MS m/z 774.1, 776.1 (M + 1 and M + 3), 792.0, 794.0 (M + H2O + 1 and M +H2O + 3). 1H NMR 400 M Hz (CD3CN-d3): δ 7.91(d, 1H), 7.76(d, 1H),7.62(dd, 1H), 7.52(dd, 1H), 7.4-7.5(m, 1H), 7.3(m, 2H), 7.2(m, 2H),6.96(d, 1H), 6.89(d, 1H), 6.77(d, 1H), 5.47(m,1H), 4.54(dd, 2H), 4.40(t,1H), 4.21(s, 1H), 4.15(m, 1H), 3.64(dd, 1H), 3.41(d, 1H), 2.93(s, 3H),2.8-3.0(m, 2H), 2.6-2.8(m, 2H), 2.2-2.4(m, 1H), 2.0-2.2(m, 1H),1.8-2.0(m, 2H), 1.5- 1.8(m, 6H).  26

MS m/z 752.4 (M + 1), 770.4 (M + H2O + 1). 1H NMR 400 M Hz (CD3CN-d3): δ7.71(d, 1H), 7.56(d, 1H), 7.41(t, 1H), 7.33(t, 1H), 6.8-7.2(m, 10H),4.7-5.1(m, 3H), 4.1-4.3(m, 2H), 3.5-3.8(m, 2H), 3.2-3.4(m, 1H),3.0-3.1(m, 1H), 2.9-3.0(m, 1H), 2.5-2.6(m, 1H), 2.3-2.5(m, 1H),1.8-2.0(m, 2H), 1.2- 1.7(m, 13H), 0.9-1.0(m, 4H), 0.6-0.8(m, 2H).  27

MS m/z 744.4 (M + 1), 762.5 (M + H2O + 1). 1H NMR 400 M Hz (CD3CN-d3): δ7.92(d, 1H), 7.85(d, NH), 7.76(d, 1H), 7.62(t, 1H), 7.52(t, 1H),7.2-7.5(m, 5H), 6.61(d, NH), 5.22(m, 1H), 5.08(dd, 2H), 4.47(m, 1H),4.00(m, 1H), 3.60(m, 1H), 3.19(dd, 2H), 2.91(m, 2H), 1.9-2.1(m, 2H),1.5- 1.8(m, 16H), 1.3-1.5(m, 6H), 1.1-1.3(m, 6H), 0.9- 1.0(m, 4H).  28

MS m/z 806.4 (M + 1), 824.4 (M + H2O + 1).  29

MS m/z 814.3, 816.3 (M + 1 and M + 3), 832.3, 834.3 (M + H2O + 1 and M +H2O + 3).  30

MS m/z 696.3 (M + 1), 714.3 (M + H2O + 1).  31

MS m/z 758.2, 760.2 (M + 1 and M + 3), 776.2, 778.2 (M + H2O + 1 and M +H2O + 3).  32

MS m/z 731.30 (M + 1)  33

MS m/z 746.3 (M + 1). Anal (C₄₃H₄₇N₅O₇•2TFA)  34

MS m/z 838.3 (M + 1)  35

MS m/z 762.3 (M + 1)  36

MS m/z 752.3 (M + 1)  37

MS m/z 732.3 (M + 1)  38

MS m/z 718.2 (M + 1)  39

MS m/z 822.3 (M + 1)  40

MS m/z 822.4 (M + 1)  41

MS m/z 731.3 (M + 1)  42

MS m/z 857.30 (M + 1). Anal. (C₄₇H₄₆ClN₇O₇•2TFA•H2O). 400 M Hz NMR(DMSO-d6) δ 9.54(s, 1H); 7.96(d, J=5.6 Hz, 1H); 7.80(d, J=5.6 Hz, 1H);7.65(m, 1H); 7.55(m, 1H); 7.43(m, 4H); 7.36(m, 4H); 7.32(m, 2H); 7.28(m,2H); 7.23(m, 3H); 7.18(m, 3H); 7.12(m, 3H); 6.40(m, 1H); 5.51(m, 1H);5.16(d, J=8.4 Hz, 1H); 5.05(d, J=8.4 Hz, 1H); 4.39(m, 1H); 4.30(m, 1H);4.25(m, 2H); 3.91(m, 1H); 3.64(m, 1H); 3.46(m, 1H); 3.36(m, 1H); 3.13(m,1H); 2.75(m, 1H); 2.64(m, 1H); 1.95(m, 2H); 1.87(m, 3H)  43

MS m/z 836.3 (M + 1)  44

MS m/z 828.3 (M + 1)  45

MS m/z 828.3 (M + 1)  46

MS m/z 870.4 (M + 1)  47

MS m/z 696.9 (M + 1) δ ¹H NMR 7.90(d, 1H J=8.0 Hz), 7.75(d, 1H J=8.0Hz), 7.60- 7.64(m, 2H), 7.51-7.55(m, 1H), 7.36-7.43(m, 1H), 7.24-7.29(m,6H), 5.46-5.49(m, 1H), 4.46-4.50(m, 1H), 4.15-4.18(m, 1H), 4.02(s, 1H),3.49-3.52(m, 1H), 3.34-3.37(m, 1H), 3.10-3.20(m, 1H), 2.98-3.05 (m, 2H),2.92(s, 3H), 2.83-2.90(m, 2H), 2.70-2.77 (m, 1H), 2.28-2.34(m, 1H),2.16-2.21(m, 1H), 1.89- 1.96(m, 3H), 1.67-1.70(m, 10H), 1.46-1.48(m,2H), 1.17-1.27(m, 4H), 0.88-0.97(m, 3H).  48

MS m/z 730.5 (M + 1)  49

MS m/z 822.4 (M + 1)  50

MS m/z 716.45 (M + 1) Anal. (C₄₀H₅₃N₅O₇•2TFA•H₂O).  51

MS m/z 730.5 (M + 1) Anal. (C₄₁H₅₅N₅O₇•2TFA•H₂O).  52

MS m/z 744.5 (M + 1)  53

MS m/z 772.4 (M + 1)  54

MS m/z 827.3 (M + 1)  55

MS m/z 744.50 (M + 1)  56

MS m/z 764.4 (M + 1)  57

MS m/z 764.4 (M + 1)  58

MS m/z 752.5 (M + 1)  59

MS m/z 820.5 (M + 1)  60

MS m/z 784.5 (M + 1)  61

MS m/z 826.5 (M + 1). NMR ((CD₃)₂CO) δ 7.97(m, 2H); 7.81(m, 3H); 7.66(m,4H); 7.56(m, 2H); 7.21(m, 1H); 6.55(m, 1H); 5.58(m, 1H); 4.57(m, 1H);4.44(m, 1H); 4.07(m, 1H); 3.97(m, 2H); 3.70(m, 1H); 3.68(m, 1H); 3.56(m,2H); 3.20(m, 2H); 2.78(m, 2H); 2.16(m, 2H); 1.71(m, 17H); 1.29(m, 7H);0.93(m, 3H).  62

MS m/z 772.3 (M + 1) NMR ((CD₃)₂CO) δ 7.95(m, 1H); 7.81(m, 1H); 7.66(m,1H); 7.56(m, 1H); 7.40(m, 4H); 7.24(m, 6H); 7.01(m, 1H); 5.44(m, 1H);4.51(m, 5H); 4.29(m, 2H); 3.81(m, 4H); 2.75(m, 2H); 2.55(m, 3H); 2.31(m,1H); 2.16(m, 1H); 1.87(m, 4H); 1.52(m, 1H); 1.36(m, 5H).  63

MS m/z 596.2 (M + 1)  64

MS m/z 660.2 (M + 1)  65

MS m/z 736.3 (M + 1)  66

MS m/z 704.2 (M + 1)  67

MS m/z 708.3 (M + 1)  68

MS m/z 708.3 (M + 1)  69

MS m/z 800.2 (M + 1) NMR ((CD₃)₂CO) δ 10.49(m, 1H); 8.02(d, J=8.0 Hz,1H); 7.86(m, 2H); 7.67(m, 1H); 7.61(m, 1H); 7.49(m, 1H); 7.32(m, 7H);7.29(m, 5H); 7.21(m, 2H); 6.49(m, 1H); 5.89(m, 1H); 4.54(m, 1H); 4.43(m,2H); 4.17(m, 2H); 3.54(m, 2H); 3.37(m, 2H); 3.08(m, 1H); 2.86(m, 1H);2.79(s, 3H); 2.74(m, 1H); 2.31(m, 1H); 1.99(m, 1H); 1.86(m, 1H).  70

MS m/z 714.3 (M + 1)  71

MS m/z 688.3 (M + 1)  72

MS m/z 752.2 (M + 1) NMR ((CD₃)₂CO) δ 8.47(m, 1H); 8.20(d, J=8.0 Hz,1H); 7.97(d, J=8.0 Hz, 1H); 7.82(d, J=8.0 Hz, 1H); 7.67(m, 2H); 7.57(m,2H); 7.36(m, 5H); 7.27(m, 3H) 7.01(m, 1H) 6.62(m, 1H); 5.62(m, 1H);4.68(m, 1H); 4.52(m, 3H); 4.30(m, 1H); 3.68(m, 1H); 3.53(m, 2H); 3.38(m,2H); 2.92(s, 3H); 2.82(m, 1H); 2.79(m, 1H); 2.39(m, 1H); 2.27(m, 1H);1.94(m, 1H).  73

MS m/z 700.3 (M + 1) NMR ((CD₃)₂CO) δ 8.00(m, 2H); 7.82(m, 1H); 7.68(m,1H); 7.58(m, 1H); 7.44(m, 6H); 7.35(m, 4H); 7.22(m, 6H); 5.78(m, 1H);4.98(m, 2H); 4.65(m, 1H); 4.47(m, 2H); 4.20(m, 2H); 3.60(m, 1H); 3.47(m,2H); 3.21(m, 1H); 2.87(m, 1H); 2.74(s, 3H); 2.55(m, 2H); 2.32(m, 1H);1.92(m, 2H).  74

MS m/z 772.2 (M + 1) NMR ((CD₃)₂CO) δ 8.30(m, 1H); 8.02(m, 1H); 7.85(m,1H); 7.73(m, 2H); 7.59(m, 1H); 7.34(m, 6H); 7.27(m, 5H); 7.23(m, 3H);5.83(m, 1H); 5.05(m, 2H); 4.81(m, 1H); 4.49(m, 2H); 4.25(m, 3H); 3.50(m,3H); 3.27(m, 1H); 2.91(m, 1H); 2.79(s, 3H); 2.79(m, 1H); 2.75(m, 1H);2.39(m, 1H); 1.95(m, 1H).  75

MS m/z 772.2 (M + 1)  76

MS m/z 736.2 (M + 1) NMR ((CD₃)₂CO) δ 11.10(m, 1H); 8.36(m, 1H); 8.20(m,1H); 8.01(m, 1H); 7.84(m, 1H); 7.68(m, 2H); 7.54(m, 2H); 7.30(m, 5H);7.23(m, 2H); 7.21(m, 2H); 7.15(m, 3H); 5.92(m, 1H); 4.52(m, 6H); 4.11(m,1H); 3.78(m, 3H); 3.09(m, 1H); 2.80(m, 3H); 2.33(m, 4H).  77

MS m/z 722.2 (M + 1) NMR ((CD₃)₂CO) δ 11.36(m, 1H); 8.60(m, 1H); 8.31(m,1H); 7.98(m, 1H); 7.91(m, 2H); 7.83(m, 1H); 7.68(m, 3H); 7.58(m, 1H);7.39(m, 1H); 7.31(m, 1H); 7.23(m, 3H); 7.17(m, 3H); 5.77(m, 1H); 4.69(m,1H); 4.52(m, 2H); 4.39(m, 2H); 3.93(m, 1H); 3.81(m, 1H); 3.65(m, 1H);3.30(m, 1H); 2.96(m, 1H); 2.78(m, 5H); 2.41(m, 2H); 2.01(m, 1H).  78

MS m/z 721.3 (M + 1)  79

MS m/z 721.3 (M + 1)  80

MS m/z 724.4 (M + 1). Anal. (C₄₁H₄₉N₅O₇S•2H₂O•TFA). 400 M Hz NMR((CD₃)₂CO) δ 7.95(m, 1H); 7.84(d, J=8.0 Hz, 2H); 7.67(d, J=8.0 Hz, 2H);7.52(m, 2H); 7.41(m, 2H); 7.39(m, 3H); 7.11(m, 5H); 6.45(d, J=4.6 Hz,1H); 5.28(m, 1H); 4.93(m, 1H); 4.49(m, 1H); 4.38(m, 2H); 4.27(m, 1H);4.14(m, 1H); 3.67(m, 3H); 3.48(m, 1H); 2.57(m, 3H); 2.42(m, 4H); 2.15(m,1H); 2.04(m, 2H); 1.90(m, 1H); 1.70(m, 3H); 1.55(m, 3H); 1.42(m, 3H). 81

MS m/z 710.4 (M + 1) (C₄₀H₄₇N₅O₇S•2H₂O•TFA). NMR ((CD₃)₂CO) δ 8.10(m,1H); 7.96(d, J=8 Hz, 2H); 7.81(d, J=8.4 Hz, 2H); 7.65(m, 2H); 7.56(m,2H); 7.34(m, 3H); 7.27(m, 5H); 7.20(m, 2H); 6.68(m, 1H); 5.44(m, 1H);4.63(m, 1H); 4.51(m, 2H); 4.44(m, 1H); 4.28(m, 1H); 3.87(m, 4H); 3.64(m,1H); 2.73(m, 2H); 2.30(m, 2H); 2.18(m, 1H); 1.85(m, 3H); 1.70(m, 1H);1.12(m, 1H); 0.52(m, 2H); 0.29(m, 2H).  82

MS m/z 634.35 (M + 1) NMR ((CD₃)₂CO) δ 8.73(m, 1H); 8.25(m, 1H); 7.95(m,1H); 7.81(m, 1H); 7.66(m, 1H); 7.36(m, 5H); 7.29(m, 1H); 5.51(m, 1H);4.67(m, 1H); 4.58(m, 4H); 4.36(m, 1H); 3.99(m, 1H); 3.60(m, 6H); 2.59(m,3H); 2.32(m, 1H); 2.17(m, 1H); 1.85(m, 4H); 1.73(m, 3H); 1.52(m, 1H);1.37(m, 4H).  83

MS m/z 732.9 (M + 1) δ ¹H NMR 8.68(s, 1H), 7.92(d, 1H J=8.0 Hz), 7.76(d,1H J= 8.0 Hz), 7.52-7.65(m, 3H), 7.40-7.44(m, 1H), 7.36 (d, 1H J=8.0Hz), 7.25-7.35(m, 3H), 6.93-9.95(m, 5H), 5.52-5.56(m, 1H), 5.07-5.09(m,1H), 4.51-4.56 (m, 1H), 4.12-4.22(m, 5H), 3.89(d, 1H J=11.2 Hz),3.77-3.81(m, 1H), 3.23-3.26(m, 2H), 2.97-3.02(m, 2H), 2.34-2.40(, 1H),2.17-2.26(m, 1H), 1.52-1.83 (m,), 1.16-1.27(m, 4H), 0.91-0.96(m, 3H). 84

MS m/z 697.9 (M + 1) δ ¹H NMR 7.85(d, 1H J=8.0 Hz), 7.68(d, 1H J=8.0Hz), 7.34- 7.35(m, 3H), 7.13-7.22(m, 4H), 6.39-6.42(m, 1H), 5.62(s, 1H),5.44-5.50(m, 1H), 4.30-4.38(m, 1H), 4.08-4.10(m, 1H), 3.94(s, 1H),3.57-3.61(m, 1H), 3.37-3.40(m, 1H), 3.01-3.13(m, 2H), 2.69-2.76(m, 1H),2.54-2.60(m, 1H), 2.42(s, 1H), 1.68-1.69(m, 2H), 1.55-1.60(m, 8H),1.35-1.39(m, 2H), 1.19(s, 1H), 1.07-1.14(m, 3H), 0.78-0.87(m, 2H).  85

MS m/z 645.2 (M + 1) δ ¹H NMR 9.16(s, 1H), 8.04(d, 1H J=8.0 Hz), 7.94(d,1H J= 8.0 Hz), 7.78-7.83(m, 2H), 7.63-7.67(m, 2H), 7.47- 7.57(m, 3H),7.17-7.14(m, 8H), 6.27(s, 2H), 5.48- 5.54(m, 1H), 4.51-4.57(m, 3H),4.42-4.46(m, 2H), 4.31-4.36(m, 1H), 3.83-3.92(m, 1H), 3.73-3.789 (m,1H), 2.33-2.39(m, 1H), 2.04-2.19(m, 2H), 1.68- 1.89(m, 3H), 1.49-1.65(m,2H)  86

MS m/z 718.4 (M + 1)  87

MS m/z 752.4 (M + 1) ¹H NMR (Acetone-d6, 400 M Hz) δ 7.99-7.92(1H, m),7.83-7.79(1H, m), 7.68-7.62(1H, m), 7.58-7.52 (1H, m), 7.46-7.12(10H,m), 5.24-5.01(3H, m), (4.54-4.35(2H, m), 4.20-4.02(1H, m), 3.87-3.51(2H,m), 3.32-3.13(2H, m), 2.90-2.65(2H, m), 2.64-2.43 (2H, m), 2.00-1.93(1H,m), 1.90-1.42(14H, m), 1.32-0.85(6H, m).  88

MS m/z 731.4 (M + 1)  89

MS m/z 775.3 (M + 1)  90

MS m/z 807.4 (M + 1)  91

MS m/z 779.4 (M + 1)  92

MS m/z 779.4 (M + 1)  93

MS m/z 792.4 (M + 1)  94

MS m/z 792.4 (M + 1)  95

MS m/z 771.4 (M + 1)  96

MS m/z 785.4 (M + 1)  97

MS m/z 786.1, 788.0 (M + 1 and M + 3), 804.1, 806.2 (M + H2O + 1 and M +H2O + 3).  98

MS m/z 758.2 (M + 1), 776.2 (M + H2O + 1). 1H NMR 400 M Hz (CD3CN-d3): δ7.93(d, 1H), 7.76(d, 1H), 7.62(t, 1H), 7.53(t, 1H), 7.15-7.35(m, 1H),7.05(m, 2H), 5.78(m, 1H), 5.62(m, 1H), 4.3- 4.5(m, 4H), 4.0-4.1(m, 2H),3.51(dd, 1H), 3.33(m, 2H), 3.0(dd, 1H), 2.76(s, 3H), 2.6-2.8(m, 3H),2.15- 2.25(m, 1H), 1.75-1.90(m, 3H).  99

MS m/z 890.3 Cl³⁵(M + 1), 892.3 Cl³⁷(M + 1). NMR ((CD₃)₂CO) δ 8.05(m,1H); 7.98(m, 2H); 7.90(m, 1H); 7.82(m, 2H); 7.66(m, 2H); 7.55(m, 2H);7.39(m, 2H); 7.34(m, 4H); 7.03(m, 2H); 6.87(m, 1H); 5.77(m, 1H); 5.60(m,1H); 5.17(m, 2H); 5.10(m, 2H); 4.57(m, 1H); 4.50(m, 2H); 4.35(m, 2H);4.17(m, 1H); 4.04(m, 1H); 3.72(m, 6H); 2.81(m, 2H); 2.72(m, 2H); 2.14(m,2H); 1.99(m, 1H). 100

MS m/z 830.2 (M + 1), 848.2 (M + H2O + 1). 1H NMR 400 M Hz (CD₃CN-d3,diasteroisomers): δ 9.03, 8.66(2s, 1H), 7.94(dd, 1H), 7.78(dd, 1H),7.64(m, 1H), 7.61(m, 1H), 7.2-7.4(m, 1H), 6.9(ddd, 1H), 6.06, 5.71(2d,1H), 5.7, 5.6(2m, 1H), 4.2-4.4(m, 3H), 4.0-4.1(m, 2H), 3.88, 3.86(2s,3H), 3.4-3.6(m, 3H), 3.2-3.4(ddd, 1H), 2.9-3.1(dt, 1H), 2.7-2.9(m, 1H),2.8, 2.7(2s, 3H), 2.6-2.7(m, 1H), 2.1-2.3(m, 1H), 1.9-2.0(m, 1H),1.7-1.9(m, 2H). 101

MS m/z 750.3 (M + 1) 102

MS m/z 770.2 (M + 1) 103

MS m/z 808.4 (M + 1) 104

MS m/z 870.4 (M + 1) 105

MS m/z 862.4 (M + 1) NMR ((CD₃)₂CO) δ 11.20(m, 1H); 8.07(m, 1H); 7.98(m,2H); 7.80(m, 3H); 7.67(m, 2H); 7.56(m, 2H); 7.44(m, 1H); 7.40(m, 2H);7.04(m, 2H); 6.86(m, 1H); 5.60(m, 1H); 4.51(m, 1H); 4.41(m, 1H); 4.35(m,1H); 3.95(m, 1H); 3.66(m, 3H); 3.35(m, 3H); 3.07(m, 4H); 2.84(m, 1H);2.74(m, 1H); 1.39(m, 2H); 1.17(m, 3H); 0.87(m, 3H). 106

MS m/z 794.3 (M + 1); ¹H NMR (CD₃CN, 600 M Hz) δ 8.75(1H, br s),8.46(1H, br s), 7.94(1H, d, J=7.8 Hz), 7.78(1H, d, J=8.4 Hz),7.65-7.61(2H, m), 7.54(1H, t, J=7.2 Hz), 7.41-7.18(11H, m), 6.89(1H, d,J=7.2 Hz), 5.32-5.30(1H, m), 5.15 (1H, d, J=12.6 Hz), 5.10(1H, d, J=12.6Hz), 4.49- 4.33(3H, m), 4.22(1H, br s), 3.77(1H, dd, J=11.4, 4.8 Hz),3.53(1H, d, J=11.4 Hz), 2.97(1H, br s), 2.81-2.73(2H, m), 2.69-2.57(5H,m), 2.29-2.20(1H, m), 2.13-1.99(2H, m), 1.95-1.89(2H, m), 1.82-1.64 (2H,m), 1.63-1.46(2H, m). 107

MS m/z 820.4 (M + 1) 108

MS m/z 794.3 (M + 1) 109

MS m/z 762.5 (M + 1) 110

MS m/z 794.3 (M + 1) 111

MS m/z 814.3 (M + 1) 112

MS m/z 742.2 (M + 1) 113

MS m/z 742.6 (M + 1) 114

MS m/z 862.4 (M + 1). 1H NMR 400 M Hz (CD3CN-d3, diasteroisomers): δ7.90(t, 1H), 7.74(t, 1H), 7.61(t, 1H), 7.52(t, 1H), 7.1- 7.5(m, 14H),5.3-5.5(m, 1H), 5.0-5.2(m, 2H), 4.1- 4.5(m, 5H), 3.4-3.8(m, 4H),2.8-3.0(m, 2H), 2.6- 2.8(m, 2H), 2.1-2.3(m, 1H), 2.0-2.1(m, 1H), 1.7-1.9(m, 2H), 1.5-1.7(m, 2H), 1.3-1.5(m, 1H), 0.9- 1.3(m, 5H). 115

MS m/z 674.3 (M + 1) 116

MS m/z 714.3 (M + 1) 117

MS m/z 716.4 (M + 1) 118

MS m/z 738.4 (M + 1) 119

MS m/z 770.3 (M + 1) 120

MS m/z 728.4 (M + 1) 121

MS m/z 730.4 (M + 1) 122

MS m/z 731.4 (M + 1) 123

MS m/z 784.3 (M + 1) 124

MS m/z 736.2 (M + 1) 125

MS m/z 728.2 (M + 1) 126

MS m/z 786.4 (M + 1) 127

MS m/z 780.2 (M + 1) 128

MS m/z 770.2 (M + 1) 129

MS m/z 729.3 (M + 1); ¹H NMR (CD₃CN, 400 M Hz) δ 9.58(1H, br s),8.31(1H, d, J=8.8 Hz), 7.93(1H, d, J=8.0 Hz), 7.78(1H, t, J=7.2 Hz),7.64(1H, t, J=8.0 Hz), 7.55(1H, t, J=7.6 Hz), 7.40-7.33(4H, m), 6.98(1H,d, J=8.8 Hz), 5.65- 5.55(1H, m), 5.41-5.30(1H, m), 4.60(1H, t, J=8.4Hz), 4.55-4.39(2H, m), 3.89(1H, d, J=11.6 Hz), 3.82(1H, dd, J=11.6, 4.0Hz), 3.28-3.16(1H, m), 3.15-3.00(2H, m), 2.98(3H, s), 2.92-2.73(2H, m),2.67-2.41(6H, m), 2.40-2.29(2H, m), 2.28-2.12(2H, m), 1.88-1.62(6H, m).130

MS m/z 714.4 (M + 1) 131

MS m/z 805.4 (M + 1) 132

MS m/z 785.3 (M + 1) 133

MS m/z 728.4 (M + 1) 134

MS m/z 729.4 (M + 1) 135

MS m/z 756.4 (M + 1) 136

MS m/z 756.4 (M + 1) 137

MS m/z 764.4 (M + 1) 138

MS m/z 822.2 (M + 1); ¹H NMR (CDCl₃, 400 M Hz) δ 7.86(1H, dd,J=6.8 Hz),7.62 (1H, d, J=8.0 Hz), 7.54(1H, t, J=8.0 Hz), 7.45 (1H, t, J=7.2 Hz),7.36-7.08(12H, m), 6.84(3H, br s), 6.11-6.09(1H, m), 5.55-5.46(1H, m),5.16-4.93 (2H, m), 4.66-4.61(1H, m), 4.21-4.06(3H, m), 3.76- 3.60(2H,m), 3.25-2.96(2H, m), 2.82-2.54(6H, m), 2.52-2.49(2H, m), 2.07-1.88(2H,m). 139

MS m/z 824.4 (M + 1); ¹H NMR (CDCl₃, 400 M Hz) δ 7.84(1H, dd, J=7.2 Hz),7.73-7.71(1H, m), 7.60- 7.59(1H, m), 7.50(1H, t, J=7.8 Hz), 7.46-7.41(1H, m), 7.31-7.05(12H, m), 6.87-6.73(2H, m), 5.56-5.53(1H, m),5.11-4.98(1H, m), 5.01-4.98(1H, m), 4.66-4.53(1H, m), 4.20-3.97(2H, m),3.89-3.80 (1H, m), 3.76-3.50(6H, m), 2.71-2.56(2H, m), 2.31- 2.12(2H,m), 2.11-1.73(4H, m), 1.70-1.31(6H, m). 140

MS m/z 828.3 (M + 1); Anal. Calcd. for C₄₃H₄₉Cl₂N₅O₉S (1 HCl + 1 H₂O):C, 58.50; H, 5.59; N, 7.93; Found: C, 58.08; H, 5.10; N, 7.79; ¹H NMR(CD₃CN, 600 M Hz) δ 8.06(1H, d, J=7.2 Hz), 7.90 (1H, d, J=8.4 Hz,7.74(3H, d, J=8.4 Hz), 7.60 (1H, t, J=7.8 Hz), 7.50(1H, t, J=7.2 Hz),7.40- 7.33(6H, m), 7.29-7.24(3H, m), 7.21-7.16(2H, m), 6.99 (1H, d,J=7.2 Hz), 5.38-5.34(1H, m), 5.10-5.08 (3H, m), 4.69-4.67(2H, m),4.35-4.29(4H, m), 4.22 (1H, br s), 4.09-4.06(1H, t, J=8.4 Hz), 3.97-3.93(1H, m), 3.82(2H, m), 2.91-2.90(2H, m), 2.76-2.72 (1H, m), 2.64-2.59(1H,m), 2.56-2.50(1H, m), 2.30- 2.28(2H, m), 1.79-1.73(2H, m), 1.61-1.57(2H,m), 1.53-1.50(1H, m). 141

MS m/z 833.3 (M + 1) 142

MS m/z 777.2 (M + 1) 143

MS m/z 779.3 (M + 1) 144

MS m/z 792.3 (M + 1) 145

MS m/z 808.2 (M + 1) 146

MS m/z 822.3 (M + 1) 147

MS m/z 844.4 (M − H₂O + 1) 148

MS m/z 794.3 (M + 1) 149

MS m/z 793.3 (M + 1) 150

MS m/z 918.3 (M + 1) 151

MS m/z 764.3 (M + 1) 152

MS m/z 872.3 (M + 1) 258

MS m/z 767.4 (M + 1)

EXAMPLE 153

In Example 153, the reagents and conditions are: (a) Cbz-OSu, Et₃N, THF,water; (b) Cbz-D-homoPhe-OH, HATU, DIEA, DCM; (c)p-nitrophenylchloroformate, pyridine, DCM; (d) piperidine, DCM; (e)LiOH.H₂O, dioxane/water (50:50 vol); (f) Reference compound 1-E, HATU,DIEA, DCM; (g) Dess-Martin periodinane, DCM; (h) TFA, DCM.

153-B: D-Homophenylalanine (3.22 g, 18.0 mmol) andN-(Benzyloxycarbonyloxy)-succinimide (Cbz-OSu) (4.49 g, 18.0 mmol) areadded to a round bottomed flask containing THF (60 mL) and water (20mL). The mixture is stirred at room temperature and Et₃N (10.1 mL, 72.0mmol) is added, and the reaction is stirred overnight at roomtemperature. The clear solution is diluted with EtOAc (200 mL); washedwith 1N HCl (3×100 mL) and brine (1×100 mL); and dried with MgSO₄.Solvent is evaporated in vacuo to afford the desired product as a whitesolid which is used without further purification.

153-D: H-Hyp-OMe.HCl (3.19 g, 17.55 mmol) is added to a stirringsolution of Cbz-D-homophenylalanine (5 g, 15.95 mmol), HATU (7.28 g,19.14 mmol), and DIEA (6.112 mL, 35.1 mmol) in DCM (100 mL). After 3hours of stirring, the mixture is washed three times with aqueous NaHSO₄1M, saturated aqueous NaHCO₃ and brine respectively. The organic phaseis dried (MgSO₄), concentrated in vacuo, and the residue is purified bychromatography on silica gel with a gradient of Ethyl Acetate/Hexanes (0to 100%) to give 1-C as a brown solid. MS m/z 441.2 (M+1), ¹H NMR(Acetone-d₆, 400 MHz) δ. 7.43-7.17 (10H, m), 6.75 (1H, d, J=8.8 Hz),5.24-5.11 (2H, m), 4.60-4.46 (1H, m), 3.81-3.55 (5H, m), 2.81-2.64 (2H,m), 2.32-2.25 (1H, m), 2.14-1.98 (3H, m).

153-E: 4-Nitrophenylchloroformate (1.514 g, 7.51 mmol) is added to asolution of 2-C (3 g, 6.83 mmol) and pyridine (663 μl, 8.19 mmol) in DCM(100 mL). The reaction mixture is stirred overnight. The mixture iswashed with three portions of NaHSO₄ 1M and two portions of brine, dried(MgSO₄) and concentrated in vacuo to give the compound 1-D as a yellowoil. MS m/z 606.2 (M+1), ¹H NMR (Acetone-d₆, 400 MHz) δ 8.34 (2H, d,J=9.2 Hz), 7.55 (2H, d, J=9.2 Hz), 7.45-7.16 (10H, m), 6.63 (1H, d,J=9.2 Hz), 5.51-5.48 (1H, m), 5.27-5.05 (2H, m), 4.56-4.50 (2H, m),4.15-3.84 (2H, m), 3.66 (3H, s), 2.80-2.55 (3H, m), 2.37-2.26 (1H, m),2.07-1.91 (2H, m).

153-F: Piperidine (320 mg, 3.76 mmol) is added to a solution of 1-D (1.9g, 3.14 mmol) in DCM (100 mL), and the solution mixture is stirred atroom temperature for 3 hours. The mixture is then washed with threeportions of aqueous 1M NaHSO₄, three portions of saturated aqueousNaHCO₃ and two portions of brine. The organic layer is dried (MgSO₄),concentrated in vacuo. The residue is purified by chromatography onsilica gel (AcOEt/Hexane, 0 to 100:%) to give the compound 1-E as abrown solid: MS m/z 552.3

(M+1); ¹H NMR (Acetone-d₆, 400 MHz) δ. 7.42-7.17 (10H, m), 6.58 (1H, d,J=8.8 Hz), 5.27-5.04 (3H, m), 4.57-4.45 (1H, m), 3.97-3.62 (5H, m),3.35-3.34 (4H, m), 2.82-2.60 (2H, m), 2.49-2.36 (1H, m), 2.21-2.21) 1H,m), 2.03-1.88 (6H, m).

153-G: Lithium hydroxide (hydrate) (37 mg, 0.88 mmol) is added to 1-E(400 mg, 0.72 mmol) in solution of THF:H₂O 50:50 (20 mL) and stirredovernight. The reaction mixture is concentrated in vacuo. The residue isdiluted with ethyl acetate; washed with three portions of aqueous NaHSO₄1M and two portions of brine; dried (MgSO₄) and concentrated in vacuo togive 1-F as a white solid. MS m/z 538.3 (M+1); ¹H NMR (Acetone-d₆, 400MHz) δ. 7.45-7.13 (10H, m), 6.82 (1H, d, J=8.4 Hz), 5.31-5.07 (3H, m),4.64-4.55 (1H, m), 4.45-3.96 (1H, m), 3.88-3.75 (1H, m), 3.36-3.30 (4H,m), 2.79-2.65 (2H, m), 2.65-2.16 (2H, m), 2.00-1.99 (2H, m), 1.60 (6H,m).

153-G: The compound 1-F (100 mg, 0.18 mmol) is added to a stirringsolution of Reference compound 1 (Scheme 1) (65 mg, 0.18 mmol). HATU (82mg, 0.21 mmol) and DIEA (70 μl, 0.40 mmol) in DCM (5 mL). After 3 hoursof stirring, the mixture is washed three times with aqueous NaHSO₄ 1M,saturated aqueous NaHCO₃ and brine respectively. The organic phase isdried (MgSO₄), concentrated in vacuo and the residue is purified bychromatography on silica gel with a gradient of ethyl acetate to Hexane0 to 100% to give 2-G as a yellow oil. MS m/z 869.4 (M+1); ¹H NMR(Acetone-d₆, 400 MHz) δ 7.71-7.62 (2H, m), 7.42-7.15 (12H, m), 5.22-5.00(4H, m), 4.56-4.38 (2H, m), 3.86-3.60 (2H, m), 3.33-3.30 (4H, m),3.06-3.00 (2H, m), 2.87-2.64 (2H, m), 2.39-2.10 (2H, m), 2.04-1.92 (2H,m), 1.79-1.50 (2H, m), 1.50-1.37 (19H, m).

153: Dess-Martin periodinane (70 mg, 0.16 mmol) is added to a stirredsolution of 1-H (120 mg, 0.14 mmol) and DCM (5 mL). The reaction mixtureis stirred for 1 h and treated with 20 mL quenching solution (25 g ofNa₂S₂O₃ in 100 mL saturated aqueous NaHCO₃). The resulting organic layeris washed twice with brine, dried (MgSO₄), and added to 20 mL of asolution of TFA (10 mL) in DCM (10 mL). The reaction mixture is stirredfor another hour, concentrated in vacuo, and the residue is purified byreverse phase HPLC (gradient of acetonitrile with 0.05% TFA and water:10 to 90%). After lyophilization, compound 153 is obtained as a whitesolid. MS m/z 767.7 (M+1), ¹H NMR (DMSO-d₆, 400 MHz)

8.98 (1H, d, J=6 Hz), 8.54 (1H, d, J=6 Hz), 8.00 (1H, d, J=8 Hz), 7.91(1H, d, J=8.4 Hz), 7.72-7.54 (2H, m), 7.41-7.11 (10H, m), 5.32-5.24 (1H,m), 5.13-4.98 (3H, m), 4.45 (1H, t, J=8 Hz), 4.23 (1H, dd, J=8 Hz, 14.4Hz), 3.99-3.47 (2H, m), 3.33-3.16 (4H, m), 2.87-2.67 (2H, m), 2.58-2.47(2H, m), 2.37-2.06 (2H, m), 2.01-1.90 (2H, m), 1.85-1.21 (12H, m). Anal(C₄₃H₄₇N₅O₇.1TFA.3H₂O).

EXAMPLES 154-240

Examples 154-240 are obtained by repeating the procedures described inExample 153, using appropriate Reference compounds and reagents whichwould be apparent to those skilled in the art, for example:

Example 154, using Reference compound 3;

Examples 155-157, using Reference Compound 4 andtrans-3-hydroxy-L-proline;

Example 158, using Reference Compound 1 and trans-3-hydroxy-L-proline;

Example 159, using Cbz-OSu and D-4-methoxyphenylglycine;

Example 160, using Cbz-OSu and D-cyclohexylalanine;

Example 161, using D-Pyroglutamic acid;

Example 162, using Cbz-D-Ala-OH;

Example 163, using Cbz-L-Lys(Boc)-OH;

Example 164, using Cbz-L-His(Boc)-OH;

Example 165, using Cbz-OSu and D-4-benzyloxyphenylglycine;

Example 166, using Cbz-OSu and D-3-trifluoromethylphenylalanine;

Example 167, using Cbz-OSu and D-4-trifluoromethylphenylalanine;

Example 168, using N-(iso-butyloxycarbonyloxy)-succinimide andD-homophenylalanine;

Example 169, using N-(ethyloxycarbonyloxy)-succinimide andD-homophenylalanine;

Example 170, using Reference compound 12;

Example 171, using Reference compound 13;

Example 172, using Reference compound 8;

Example 173, using N-(cyclohexyloxycarbonyloxy)-succinimide andD-3-chlorophenylalanine;

Example 174, using Reference compound 5;

Example 175, using Reference compound 5 and Reference compound 14;

Example 176, using N-p-tosylglycine and Reference compound 5;

Example 177, using N-Boc-4-hydroxypiperidine;

Example 178, using N—,N-diethylamine;

Example 179, using (±)-3-(tert-butoxycarbonylamino)-pyrrolidine;

Example 180, using 1-Boc-piperazine;

Example 181, using morpholine;

Example 182, using pyrrolidine;

Example 183, using N—,N-methylamine;

Example 184, using 1-acetyl-piperazine;

Example 185, using Cbz-D-Phe-OH;

Example 186, using N-benzylmethylamine;

Example 187, using N-methylfurfurylamine;

Example 188, using 4-phenylpiperidine;

Example 189, using 1-methanesulfonyl-piperazine;

Example 190, using 1-(2-furoyl)piperazine;

Example 191, using 1-(2-tetrahydrofuroyl)piperazine;

Example 192 is prepared, using 1-(benzoyl)piperazine;

Example 193, using 4-(tert-butoxycarbonylamino)-piperidine;

Example 194, using 1-phenyl sulfonyl piperazine;

Example 195, using 4-(aminomethyl)-1-N-Boc-piperidine;

Example 196, using 4-N-Boc-4-N-methyl-aminopiperidine;

Example 197, using 4-(2-aminoethyl)-1-Boc-piperidine;

Example 198, using Reference compound 14;

Example 199, using 4-methylsulphonylbenzylamine hydrochloride;

Example 200, using D-4-phenylphenylalanine;

Example 201, using D-3-methylphenylalanine;

Example 202, using D-3-chlorophenylalanine;

Example 203, using D-2-methylphenylalanine;

Example 204, using D-2-chlorophenylalanine;

Example 205, using 4-(trifluoromethoxy)-DL-phenylglycine;

Example 206, using 4-phenyl-DL-phenylglycine;

Example 207, using Reference compound 15;

Example 208, using 4,4-difluoropiperidine;

Example 209, using 2-thiophenemethylamine;

Example 210, using 1-(4-fluoro-benzenesulfonyl)-piperazine;

Example 211, using D-2-thienylalanine;

Example 212, using N-(cyclopropylmethyloxycarbonyloxy)-succinimide andD-homophenylalanine;

Example 213, using Reference compound 53;

Example 214, using N-4-piperidinyl-benzenesulfonamide;

Example 215, using D-2-fluorophenylalanine;

Example 216, using D-3-fluorophenylalanine;

Example 217, using D-4-fluorophenylalanine;

Example 218, using D-2-trifluoromethylphenylalanine;

Example 219, using 4-(trifluoromethyl)-DL-phenylglycine;

Example 220, using N-(cyclopentyloxycarbonyloxy)-succinimide andD-homophenylalanine;

Example 222, using Reference compound 2;

Example 223, using Reference compound 18;

Example 224, using Reference compound 3;

Example 225, using N-(cyclohexyloxycarbonyloxy)-succinimide andD-homophenylalanine;

Example 226, using N-p-tosylglycine;

Example 227, using N-(cyclopentyloxycarbonyloxy)-succinimide andD-homophenylalanine;

Example 228, using Reference compound 19;

Example 229, using Cbz-OSu and D-homocyclohexylalanine (D-homoCha-OH);

Example 230, using Reference compound 20;

Example 231, using Reference compound 7;

Example 232, using Reference compound 11;

Example 233, using Reference compound 9;

Example 234, using Reference compound 8;

Example 235, using Reference compound 2;

Example 236, using Reference compound 5;

Example 237, using Reference compound 6;

Example 238, using Reference compound 10;

Example 239, using N-(cyclohexyloxycarbonyloxy)-succinimide andD-allylglycine; and Example 240, using Reference compound 21.

EXAMPLE 241

In Example 241, the reagents and conditions are: (a) HATU, DIEA, DCM;(b) H₂ (40 psi), i-PrOH:H₂O (3:1) (c) HATU, DIEA, DCM; (d) Dess-Martinperiodinane, DCM; (e) Hoveyda-Grubbs metathesis catalyst,4-Methylene-N-Boc-piperidine, DCM, 40° C. (f) TFA, DCM.

241-A: Reference compound 54 is reacted with Reference compound 1,following methods analogous to those used in Step f of Example 153.Intermediate 241-B is obtained following hydrogenolysis conditionsanalogous to Step D in the synthesis of Reference compound 1.Dichloromethane (10 mL, 0.1 M) is added to intermediate 241-B (600 mg,1.049 mmol, 1.0 eq.), Reference compound 21 (203 mg, 1.049 mmol, 1.0eq.) and HATU (478 mg, 1.258 mmol, 1.2 eq.). DIEA (550 μL, 3.147 mmol,3.0 eq.) is added via syringe, and the reaction is stirred at roomtemperature to completion, as determined by LC/MS. Solution is dilutedwith ethyl acetate (100 mL) and extracted with 1M HCl (3×30 mL),saturated NaHCO₃ (1×30 mL) and saturated NaCl (1×30 mL). The organiclayer is dried over MgSO₄, filtered and evaporated to dryness to provideintermediate 241-C as a colorless oil. MS m/z 749.4 (M+1).

241-D: Dichloromethane (5 mL, 0.07 M) is added to 89-C (273 mg, 0.365mmol, 1.0 eq) and Dess-Martin Periodinane (309 mg, 0.73 mmol, 2.0 eq.)at room temperature under a nitrogen atmosphere. The reaction ismonitored to completion by LC/MS, diluted with ethyl acetate (50 mL),and extracted with saturated sodium thiosulfate (3×20 mL), saturatedNaHCO₃ (1×30 mL) and saturated NaCl (1×30 mL). The organic layer isdried over MgSO₄, filtered and evaporated to dryness. Automatedsilica-gel purification (0-100% ethyl acetate in hexanes) provided 89-Das a colorless oil. MS m/z 747.4 (M+1).

241-E: Anhydrous dichloromethane (2 mL, 0.03 M) is added via syringe to89-D (40 mg, 0.054 mmol, 1.0 eq.), Hoveyda-Grubbs 2^(nd) Generationmetathesis catalyst(1,3-Bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene)dichloro(o-isopropoxyphenylmethylene) ruthenium II dichloride) (10 mg,0.016 mmol, 30 mol %) under a nitrogen atmosphere.N-Boc-4-methylenepiperidine (53 μL, 0.268 mmol, 5.0 eq.) is added viasyringe, and the reaction is fitted with a reflux condenser, and heatedto 40° C. for 12 h. After the reaction is judged as complete by LC/MS,the reaction mixture is directly purified by automated silica-gelpurification (0-100% ethyl acetate in hexanes) to provide 241-E as adark green oil. MS m/z 816.4 (M-Boc+1).

241: Three mL of a 25:75 mixture of trifluoroacetic acid anddichloromethane is added to 241-E (18 mg, 0.02 mmol) and stirred at roomtemperature until judged to completion by LC/MS. The reaction mixture isconcentrated in vacuo and the residue is purified by reverse phase HPLC(gradient of Acetonitrile with 0.05% TFA and water: 10 to 90%). Afterlyophilization, Example 89 is obtained as a white solid. MS m/z 716.3.

EXAMPLES 242-243

Examples 242 and 243 are prepared, following methods analogous to thosedescribed for Example 241, using Reference compounds 55 and 56,respectively.

Example 244 is prepared, following methods analogous to those describedin Example 241, using methylenecyclopentane in step e.

EXAMPLE 245

In Example 245, the reagents and conditions are: (a) Hoveyda-Grubbsmetathesis catalyst, Methylenecyclopentane, DCM, 40° C. (b) H₂ (40 psi),i-PrOH:H₂O (3:1); (c) Dess-Martin periodinane, DCM; (d) TFA, DCM.

245-A: Anhydrous dichloromethane (2 mL, 0.03 M) is added via syringe to89-C (80 mg, 0.107 mmol, 1.0 eq.), Hoveyda-Grubbs 2^(nd) Generationmetathesis catalyst(1,3-Bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene)dichloro(o-isopropoxyphenylmethylene) ruthenium II dichloride) (14 mg,0.021 mmol, 20 mol %) under a nitrogen atmosphere. Methylenecyclopentane(57 μL, 0.543 mmol, 5.0 eq.) is added via syringe, and the reaction isfitted with a reflux condenser and heated to 40° C. for 12 hours. Afterthe reaction is judged as complete by LC/MS, the reaction mixture isdirectly purified by automated silica-gel purification (0-100% ethylacetate in hexanes) to provide 245-A as a dark green oil. MS m/z 803.4(M+1).

245-B: tert-Butanol (30 mL) and water (10 mL) is added to 93-A (69 mg,0.086 mmol, 1.0 eq.) and Pd/C (10 mg) in a Parr shaker. The Parr shakeris pressurized to 40 psi and shaken for 12 h. After the reaction isjudged as complete by LC/MS, the reaction mixture is filtered overCelite, and the solvents evaporated to provide 245-B as a light greenoil that is used in the next reaction without further purification. MSm/z 805.4 (M+1).

245-C: Dichloromethane (5 mL, 0.02 M) is added to 93-B (69 mg, 0.086mmol, 1.0 eq) and Dess-Martin periodinane (70 mg, 0.17 mmol, 2.0 eq.) atroom temperature under a nitrogen atmosphere. The reaction is monitoredto completion by LC/MS, diluted with ethyl acetate (50 mL), andextracted with saturated sodium thiosulfate (3×20 mL), saturated NaHCO₃(1×30 mL) and saturated NaCl (1×30 mL). The organic layer is dried overMgSO₄, filtered and evaporated to dryness. Automated silica-gelpurification (0-100% ethyl acetate in hexanes) provided 245-C as acolorless oil. MS m/z 703.4 (M-Boc+1).

245: Three mL of a 25:75 mixture of trifluoroacetic acid anddichloromethane is added to 93-C (15 mg, 0.019 mmol) and stirred at roomtemperature until it is judged to completion by LC/MS. The reactionmixture is concentrated in vacuo, and the residue is purified by reversephase HPLC (gradient of Acetonitrile with 0.05% TFA and water: 10 to90%). Following lyophilization, Example 245 is obtained as a whitesolid. MS m/z 703.3.

EXAMPLE 246

Example 246 is prepared from Example 245-A, following methods analogousto those described for the preparation of Example 153.

Table 2 shows compounds of Formula (2), as described in Examples153-246.

TABLE 2 Physical Data Structure ¹H NMR 400 M Hz (DMSO-d₆) and/or MS(m/z) 153

¹H NMR (DMSO-d6, 400 M Hz) δ 8.98(1H, d, J=6 Hz), 8.54(1H, d, J=6 Hz),8.00(1H, d, J= 8 Hz), 7.91(1H, d, J=8.4 Hz), 7.72-7.54 (2H, m),7.41-7.11(10H, m), 5.32-5.24(1H, m), 5.13-4.98(3H, m), 4.45(1H, t, J=8Hz), 4.23(1H, dd, J=8 Hz, 14.4 Hz), 3.99-3.47 (2H, m), 3.33-3.16(4H, m),2.87-2.67(2H, m), 2.58-2.47(2H, m), 2.37-2.06(2H, m), 2.01-1.90(2H, m),1.85-1.21(12H, m). Anal (C₄₃H₄₇N₅O₇•1TFA•3H₂O); MS m/z 767.4 (M + 1).154

MS m/z 811.7 (M + 1); Anal. Calcd. for C₄₄H₅₁F₃N₈O₉S (1 TFA salt): C,57.13; H, 5.56; N, 12.11; Found: C, 57.11; H, 5.38; N: 11.32; ¹H NMR(CD₃CN, 400 M Hz) δ 8.21 (1H, dd, J=5.6, 2.4 Hz), 8.10(1H, dd, J= 5.2,1.6 Hz), 7.82(1H, d, J=5.2 Hz), 7.67- 7.60(2H, m), 7.37-7.34(3H, m),7.31-7.26 (2H, m), 7.20-7.16(2H, m), 6.85(2H, br s), 6.50(1H, d, J=4.8Hz), 5.63-5.51(1H, m), 5.33(1H, dd, J=10.8, 3.2 Hz), 5.16-5.00(3H, m),4.57-4.51(1H, m), 4.30-4.26(1H, m), 3.89-3.70(3H, m), 3.59-3.49(1H, m),3.30- 3.12(2H, m), 2.77-2.72(1H, m), 2.65-2.59 (1H, m), 2.26-2.18(2H,m), 1.92-1.67(6H, m), 1.54-1.29(6H, m). 155

MS m/z 797.2 (M + 1) 156

MS m/z 795.3 (M + 1); ¹H NMR (CDCl₃, 400 M Hz) δ 7.86-7.77(1H, m),7.59-7.52(1H, m), 7.51-7.43(1H, m), 7.42-7.35(1H, m), 7.32- 7.01(10H,m), 5.62-5.47(1H, m), 5.36-5.19 (1H, m), 5.10-4.90(3H, m), 4.70-4.48(1H,m), 4.35-4.21(1H, m), 3.95-3.76(1H, m), 3.45-3.16(5H, m), 3.15-3.03(1H,m), 2.72- 2.51(3H, m), 2.10-2.00(3H, m), 1.95-1.82 (2H, m),1.76-1.55(2H, m), 1.53-1.28(6H, m). 157

MS m/z 781.3 (M + 1); ¹H NMR (CDCl₃, 400 M Hz) δ 7.86-7.77(1H, m),7.59-7.52(1H, m), 7.51-7.43(1H, m), 7.42-7.35(1H, m), 7.32- 7.01(10H,m), 5.62-5.47(1H, m), 5.36-5.19 (1H, m), 5.10-4.90(3H, m), 4.70-4.48(1H,m), 4.35-4.21(1H, m), 3.95-3.76(1H, m), 3.45-3.16(5H, m), 3.15-3.03(1H,m), 2.72- 2.51(3H, m), 2.10-2.00(3H, m), 1.95-1.82 (2H, m),1.76-1.55(2H, m), 1.53-1.28(4H, m). 158

MS m/z 767.4 (M + 1) ¹H NMR (CD₃CN, 400 M Hz) δ 7.17(1H, d, J=7.6 Hz),6.92(1H, d, J=8.4 Hz), 6.83(1H, t, J=8.0 Hz), 6.74(1H, t, J=7.6 Hz),6.69(10H, m), 6.18-6.10(1H, m), 4.96-4.85(1H, m), 4.50-4.21(3H, m),3.86-3.69(1H, m), 3.34-3.16(1H, m), 2.85- 2.49(5H, m), 2.35-2.02(8H, m),1.67-1.51 (1H, m), 1.50-1.40(1H, m), 1.39-1.13(2H, m), 1.11-0.90(2H, m),0.88-0.68(6H, m). 159

MS m/z 769.3 (M + 1) 160

MS m/z 759.4 (M + 1) 161

MS m/z 759.4 (M + 1) 162

MS m/z 677.4 (M + 1) 163

MS m/z 734.4 (M + 1) 164

MS m/z 743.3 (M + 1) 165

MS m/z 845.4 (M + 1) 166

MS m/z 821.4 (M + 1) 167

MS m/z 821.3 (M + 1) 168

MS m/z 733.4 (M + 1). NMR ((CD₃)₂CO) δ 7.82(m, 1H); 7.67(m, 1H); 7.52(m,1H); 7.42(m, 1H); 7.14(m, 5H); 6.87(m, 1H); 5.10(m, 1H); 4.54(m, 1H);4.24(m, 1H); 3.66(m 5H); 3.21(m, 4H); 2.43(m, 4H); 1.73(m, 5H); 1.58(m,2H); 1.38(m, 7H); 0.73(m, 6H) 169

MS m/z 705.4 (M + 1)). NMR ((CD₃)₂CO) δ 7.80(m, 1H); 7.66(m, 1H);7.56(m, 1H); 7.44 (m, 1H); 7.38(m, 1H); 7.27(m, 4H); 7.02(m, 1H);5.23(m, 1H); 4.05(m, 5H); 3.84(m, 5H); 3.67(m, 6H); 3.06(m, 4H); 2.71(m,5H); 2.32 (m, 2H); 1.90(m, 2H); 1.43(m, 7H); 1.20(m, 3H). 170

MS m/z 752.4 (M + 1) 171

MS m/z 675.4 (M + 1) 172

MS m/z 845.4 (M + 1) 173

MS m/z 823.35 (M + 1). Anal. (C₄₀H₅₁ClN₈O₇S•H₂O•2TFA) NMR ((CD₃)₂CO) δ8.27(m, 3H); 8.17(m, 1H); 7.60 (m, 3H); 7.34(m, 4H); 7.04(m, 1H);6.71(m, 1H); 5.76(m, 1H); 5.20(m, 1H); 4.69(m, 1H); 4.59(m, 1H); 4.50(m,1H); 4.04(m, 1H); 3.56 (m, 1H); 3.07(m, 3H); 2.28(m, 3H); 1.90(m, 6H);1.69(m, 2H); 1.60(m, 2H); 1.49(m, 7H); 1.34(m, 7H). 174

MS m/z 827.40 (M + 1)). Anal. (C₄₄H₅₁ClN₆O₈•H₂O•2TFA) NMR ((CD₃)₂CO) δ8.00(m, 1H); 7.92(m, 1H); 7.90(m, 1H); 7.68(m, 1H); 7.45(m, 6H); 7.35(m,4H); 5.81 (m, 1H); 5.03(m, 3H); 4.75(m, 1H); 4.59(m, 2H); 4.53(m, 1H);4.33(m, 1H); 3.88(m, 1H); 3.04(m, 4H); 2.62(m, 1H); 2.22(m, 1H); 1.87(m, 2H); 1.68(m, 4H); 1.50(m, 6H); 1.26(m, 6H). 175

MS m/z 759.4 (M + 1). Anal. (C₃₉H₄₆N₆O₈S•2TFA) NMR ((CD₃)₂CO) δ 8.00(m,2H); 7.82(m, 1H); 7.68(m, 1H); 7.58 (m, 1H); 7.44(m, 5H); 7.29(m, 5H);7.21(m, 1H); 5.81(m, 1H); 5.17(m, 1H); 4.99(m, 3H); 4.69(m, 1H); 4.15(m,1H); 3.70(m, 1H); 3.49 (m, 2H); 3.21(m, 1H); 2.76(s, 3H); 2.74(m, 1H);2.61(m, 1H); 2.33(m, 1H); 1.93(m, 2H); 1.53(m, 2H); 1.44(m, 4H). 176

MS m/z 731.3 (M + 1) NMR ((CD₃)₂CO) δ 7.98 (m, 2H); 7.83(m, 1H); 7.77(m,2H); 7.68(m, 1H); 7.49(m, 2H); 7.40(m, 3H); 7.35(m, 2H); 5.72(m, 1H);5.01(s, 2H); 4.52(m, 1H); 3.65(m, 3H); 3.49(m, 1H); 3.27(m, 1H); 3.03(m,1H); 2.60(m, 1H); 2.47(s, 3H); 1.87(m, 1H); 1.68(m, 8H). 177

MS m/z 781.4 (M + 1) 178

MS m/z 755.3 (M + 1) 179

MS m/z 768.5 (M + 1) 180

MS m/z 768.3 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 7.96-7.57(4H, m),7.43-7.05 10H, m), 5.27-4.83(4H, m), 4.60-4.35(1H, m), 4.35-4.15(1H, m),4.05-3.26(6H, m), 3.18-2.95(6H, m), 2.82-2.54(2H, m), 2.40- 2.00(4H, m),1.85-1.45(6H, m). 181

MS m/z 769.4 (M + 1) 182

MS m/z 753.4 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 8.04-7.52(4H, m),7.42-7.01 (10H, m), 5.17-4.93(4H, m), 4.55-4.41(1H, m), 4.33-4.13(1H,m), 3.95-3.50(2H, m), 3.33-2.97(6H, m), 2.85-2.59(2H, m), 2.41- 1.94(4H,m), 1.88-1.47(10H, m). 183

MS m/z 727.3 (M + 1) 184

MS m/z 810.4 (M + 1) 185

MS m/z 753.3 (M + 1) 186

MS m/z 803.4 (M + 1) 187

MS m/z 793.3 (M + 1) 188

MS m/z 843.4 (M + 1) 189

MS m/z 846.2 (M + 1) 190

MS m/z 862.4 (M + 1) 191

MS m/z 866.5 (M + 1) 192

MS m/z 872.5 (M + 1) 193

MS m/z 782.6 (M + 1) 194

MS m/z 908.4 (M + 1) 195

MS m/z 796.4 (M + 1) 196

MS m/z 796.4 (M + 1) 197

MS m/z 810.4 (M + 1), 405.7 ((M + 1)/2) 198

MS m/z 711.6 (M + 1), Anal. (C35H46N6O8S•2H₂O•2TFA), ¹H NMR (DMSO-d₆,400 M Hz) δ 8.69(1H, d, J=7.2 Hz), 7.98(1H, d, J=8 Hz), 7.89(1H, d, J=8.4 Hz), 7.70-7.48(2H, m), 7.33-7.11(5H, m), 5.35-5.24(1H, m),5.15-5.02(1H, m), 4.51- 4.38(1H, m), 4.07-3.92(1H, m), 3.87-3.57 (2H,m), 3.32-3.12(2H, m), 3.12-2.60(5H, m), 2.16-1.86(2H, m), 1.86-1.67(2H,m), 1.67- 1.18(12H, m). 199

MS m/z 867.3 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 8.05-7.43(4H, m),7.43-6.89 (14H, m), 5.15-4.93(4H, m), 4.53-4.00(2H, m), 3.79-3.33(2H,m), 3.16(3H, s), 2.83-2.50 (4H, m), 2.42-1.95(2H, m), 1.90-1.46(8H, m).200

MS m/z 829.4 (M + 1) 201

MS m/z 757.4 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 7.85-7.50(4H, m),7.37-6.85(9H, m), 5.20-4.85(4H, m), 4.50-4.20(2H, m), 3.95-3.65(2H, m),3.30-3.20(4H, m), 2.85- 2.65(4H, m), 2.25-2.20(3H, s), 2.20-1.90 (2H,m), 1.60-1.27(12H, m). 202

MS m/z 787.4 (M + 1) 203

MS m/z 767.4 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 8.04-7.90(2H, m),7.80-7.56(2H, m), 7.38-7.00(9H, m), 5.12-4.82(4H, m), 4.60-4.32(2H, m),3.97-3.60(2H, m), 3.36- 3.18(4H, m), 2.97-2.83(1H, m), 2.60-2.54 (1H,m), 2.30-1.75(5H, m), 1.60-1.31(12H, m). 204

MS m/z 787.4 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 8.04-7.90(2H, m),7.80-7.54(2H, m), 7.43-7.12(9H, m), 5.37-5.20(1H, m), 5.15-4.83(3H, m),4.65-4.45(2H, m), 3.85- 3.65(2H, m), 3.40-3.20(4H, m), 3.15-2.70 (4H,m), 2.30-1.94(2H, m), 1.75-1.30(12H, m). 205

MS m/z 823.3 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 7.70-7.54(2H, m),7.52-7.39(2H, m), 7.39-7.23(5H, m), 7.19-7.03(2H, m), 7.02-6.89(2H, m),5.62-5.55(1H, m), 5.20- 4.93(4H, m), 4.46-4.23(1H, m), 3.90-3.68 (2H,m), 3.46-3.17(4H, m), 2.87-2.67(2H, m), 2.38-1.82(2H, m), 1.67-1.34(12H,m). 206

MS m/z 815.4 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 7.77-7.58(4H, m),7.58-7.40(4H, m), 7.40-7.24(8H, m), 5.60-5.48(1H, m), 5.28-5.13(1H, m),5.13-4.94(3H, m), 4.50- 4.30(1H, m), 3.90-3.75(2H, m), 3.44-3.20 (4H,m), 2.92-2.70(2H, m), 2.36-1.90(2H, m), 1.66-1.27(12H, m). 207

MS m/z 831.4 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 7.76-7.50(4H, m),7.42-7.25(5H, m), 5.20-4.95(4H, m), 4.87-4.72(1H, m), 4.55-4.44(1H, m),3.85-3.45(6H, m), 3.40- 3.20(4H, m), 2.85-2.70(2H, m), 2.40-1.95 (2H,m), 1.65-1.30(12H, m). 208

MS m/z 803.4 (M + 1) 209

MS m/z 795.3 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 7.87-7.63(2H, m),7.56-7.30(2H, m), 7.22-6.84(13H, m), 5.01-4.70(4H, m), 4.30-4.18(1H, m),4.18-4.07(2H, m), 4.07- 3.82(1H, m), 3.50-3.16(2H, m), 2.67-2.51 (2H,m), 2.51-2.30(2H, m), 2.16-1.70(2H, m), 1.65-1.19(8H, m). 210

MS m/z 926.4 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 8.04-7.06(18H, m),5.17-4.94 (4H, m), 4.50-4.33(1H, m), 4.33-4.10(1H, m), 3.91-3.54(2H, m),3.54-3.25(8H, m), 2.97-2.50(4H, m), 2.33-1.87(2H, m), 1.84- 1.40(8H, m).211

MS m/z 759.3 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 7.92-7.57(4H, m),7.35-7.20(5H, m), 7.05-6.81(3H, m), 5.20-4.90(4H, m), 4.51-4.20(2H, m),3.90-3.43(2H, m), 3.35- 3.20(4H, m), 3.20-2.95(2H, m), 2.83-2.70 (2H,m), 2.33-2.00(2H, m), 1.70-1.28(12H, m). 212

MS m/z 731.4 (M + 1) 213

MS m/z 922.4 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 7.79-7.53(4H, m),7.40-6.85 (15H, m), 5.20-4.90(4H, m), 4.50-4.33(1H, m), 4.33-4.15(1H,m), 4.04-3.55(2H, m), 3.50-3.08(8H, m), 2.86-2.60(4H, m), 2.38- 1.96(2H,m), 1.88-1.34(10H, m). 214

MS m/z 922.4 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 8.01-7.45(9H, m),7.41-7.04 (10H, m), 5.15-4.87(4H, m), 4.52-4.32(1H, m), 4.27-4.13(1H,m), 3.80-3.51(2H, m), 3.33-3.05(5H, m), 2.94-2.57(4H, m), 2.33- 1.92(2H,m), 1.85-1.64(2H, m), 1.64-1.12 (10H, m). 215

MS m/z 771.4 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 8.93(1H, d, J=5.6Hz), 8.65 (1H, d, J=6.4 Hz), 8.01(1H, d, J=8 Hz), 7.91(1H, d, J=8.4 Hz),7.65(1H, t, J=8 Hz), 7.56(1H, t, J=7.6 Hz), 7.31-7.02(9H, m),5.37-5.25(1H, m), 5.14-4.82(3H, m), 4.60-4.43(2H, m), 3.87-3.37(2H, m),3.35- 3.14(4H, m), 3.01-2.50(4H, m), 2.24-1.91 (2H, m), 1.75-1.18(12H,m). 216

MS m/z 771.4 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 8.05-7.86(2H, m),7.75-7.50(2H, m), 7.40-6.92(9H, m), 5.40-5.22(1H, m), 5.15-4.80(3H, m),4.55-4.30(2H, m), 3.88- 3.40(2H, m), 3.35-3.28(4H, m), 2.95-2.53 (4H,m), 2.30-1.90(2H, m), 1.80-1.28(12H, m). 217

MS m/z 771.4 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 8.07-7.50(4H, m),7.38-6.86(9H, m), 5.46-5.24(1H, m), 5.12-4.82(4H, m), 4.53-4.30(2H, m),4.23-3.93(1H, m), 3.90- 3.74(2H, m), 3.51-3.36(1H, m), 3.36-3.12 (4H,m), 2.90-2.62(4H, m), 2.30-1.87(2H, m), 1.72-1.40(12H, m). 218

MS m/z 821.4 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 8.01-7.35(4H, m),7.35-6.85(9H, m), 5.19-4.80(4H, m), 4.62-4.36(2H, m), 3.87-3.38(2H, m),3.38-3.19(4H, m), 3.19- 3.05(1H, m), 3.05-2.87(1H, m), 2.87-2.70 (2H,m), 2.30-1.93(2H, m), 1.63-1.30(12H, m). 219

MS m/z 807.3 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 8.06-7.85(2H, m),7.82-7.47(2H, m), 7.42-7.19(5H, m), 5.60-5.46(1H, m), 5.13-4.85(3H, m),4.51-4.31(1H, m), 4.31- 4.17(1H, m), 3.87-3.40(2H, m), 3.35-2.85 (4H,m), 2.85-2.64(2H, m), 2.31-1.84(2H, m), 1.60-1.07(12H, m). 220

MS m/z 886.4 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 8.03-7.41(6H, m),7.37-7.04(8H, m), 5.14-4.83(3H, m), 4.48-4.33(1H, m), 4.23-4.08(1H, m),4.02-3.56(2H, m), 3.44- 3.23(8H, m), 2.96-2.51(4H, m), 2.30-1.92 (2H,m), 1.85-1.35(16H, m). 221

MS m/z 857.5 (M + 1), ¹H NMR (DMSO-d₆, 400 M Hz) δ 7.77-7.48(4H, m),7.48-7.02 (15H, m), 5.16-4.90(4H, m), 4.54-4.20(2H, m), 4.02-3.80(2H,m), 3.90-42-3.07(4H, m), 2.94-2.65(3H, m), 2.50-2.33(4H, m), 2.33-1.80(2H, m), 1.60-1.18(12H, m). 222

MS m/z 843.4 (M + 1) 223

MS m/z 647.4 (M + 1), Anal. (C35H46N6O6•3TFA), ¹H NMR (acetone-d₆, 400 MHz) δ 8.53(1H, d, J=6.8 Hz), 8.00- 7.94(1H, m), 7.85-7.79(1H, m),8.00-7.94 (1H, m), 7.85-7.79(1H, m), 7.70-7.62 8.00- 7.94(1H, m),7.85-7.79(1H, m), 7.59-7.53 (1H, m), 7.33-7.23(5H, m), 5.40-5.25(2H, m),4.75(1H, t, J=4.75 Hz), 4.42(1H, dd, J= 7.92 Hz, 5.25 Hz), 4.00-3.80(2H,m), 3.45- 3.16(4H, m), 2.90-2.70(2H, m), 2.47-2.33 (1H, m),2.33-2.14(2H, m), 2.1-2.08(3H, m), 2.0-1.86(1H, m), 1.86-1.67(2H, m),1.60- 1.25(12H, m). 224

MS m/z 691.4 (M + 1), Anal. (C35H46N8O5S•4TFA) 225

MS m/z 759.5 (M + 1), Anal. (C41H54N6O8•2TFA), ¹H NMR (DMSO-d6, 400 MHz) δ 8.96(1H, d, J=4 Hz), 8.52(1H, d, J=5.6 Hz), 8.00(1H, d), 7.91(1H,d), 7.70-7.52(2H, m), 7.40-7.05(5H, m), 5.30- 5.24(1H, m), 5.12-5.00(2H,m), 4.50-4.38 (2H, m), 4.27-4.13(1H, m), 3.95-3.52(2H, m), 3.40-3.06(4H,m), 2.86-2.70(2H, m), 2.70-2.53(2H, m), 2.37-1.90(4H, m), 1.84-1.11(22H, m). 226

MS m/z 683.3 (M + 1), Anal. (C33H42N6O8S•2TFA), ¹H NMR (Acetone- d₆, 400M Hz) δ 7.98-7.86(1H, m), 7.75-7.66 (1H, m), 7.62-7.54(1H, m),7.54-7.46(1H, m), 7.40-7.27(4H, m), 5.51-5.40(1H, m), 5.21-5.13(1H, m),4.65-4.55(1H, m), 3.89- 3.41(4H, m), 3.37-3.20(4H, m), 2.60-2.43 (2H,m), 2.37(3H, s), 2.30-2.06(2H, m), 1.96- 1.32(12H, m). 227

¹NMR (DMSO-d6, 400 M Hz) δ 8.98(1H, d, J=6 Hz), 8.52(1H, d, J=6 Hz),8.00(1H, d, J=7.6 Hz), 7.91(1H, d, J=8.4 Hz), 7.76- 7.54(2H, m),7.36-7.12(5H, m), 5.33-5.23 (1H, m), 5.13-4.87(3H, m), 4.44(1H, t, J=8Hz), 4.18(1H, dd, J=7.2 Hz, 14 Hz), 3.94- 3.45(2H, m), 3.38-3.16(4H, m),2.88-2.70 (2H, m), 2.70-2.45(2H, m), 2.35-2.04(2H, m), 2.03-1.91(2H, m),1.85-1.24(20H, m); Anal. (C₄₀H₅₂N₆O₈•2TFA)s 228

MS m/z 717.3 (M + 1). NMR ((CD₃)₂CO) δ 8.32(m, 1H); 7.97(m, 2H); 7.82(m,2H); 7.67(m, 2H); 7.57(m, 2H); 6.31(m, 1H); 5.52(m, 1H); 5.28(m, 1H);4.74(m, 1H); 4.13(m, 1H); 3.95(m, 1H); 3.92-3.80(m, 4H); 2.97(m, 1H);2.88(m, 4H); 2.62-2.52(m, 4H); 2.44(m, 1H); 2.27-2.17(m, 1H);1.99-1.81(m, 3H); 1.76-1.59(m, 5H); 1.56-1.39(m, 6H); 1.35-1.13(m, 6H);0.96-0.87(m, 3H) 229

MS m/z 773.4 (M + 1). NMR ((CD₃)₂CO) δ 8.01(m, 1H); 7.86(m, 2H); 7.70(m,2H); 7.55(m, 2H)7.47-7.43(m, 5H); 7.19(m, 1H); 6.55(m, 1H); 5.37(m, 1H);5.16(m, 1H); 4.99(m, 3H); 4.56(m, 1H); 4.28(m, 1H); 3.88(m, 1H);3.74-3.62(m, 4H); 2.48-2.39(m, 4H); 2.25-2.11(m, 3H); 1.83-1.68(m, 3H);1.65-1.48(m, 8H); 1.45-1.33(m, 4H); 1.28- 0.99(m, 7H); 0.84-0.73(m, 2H).230

MS m/z 718.4 (M + 1) 231

MS m/z 772.4 (M + 1) 232

MS m/z 758.4 (M + 1) 233

MS m/z 766.4 (M + 1) 234

MS m/z 794.4 (M + 1) 235

MS m/z 794.4 (M + 1) 236

MS m/z 766.4 (M + 1) 237

MS m/z 779.4 (M + 1) 238

MS m/z 779.4 (M + 1) 239

MS m/z 695.4 (M + 1) 240

MS m/z 647.4 (M + 1) 241

MS m/z 716.4 (M + 1) 242

MS m/z 772.4 (M + 1) 243

MS m/z 750.4 (M + 1) 244

MS m/z 778.4 (M + 1) 245

MS m/z 703.3 (M + 1) 246

MS m/z 701.4 (M + 1)

EXAMPLE 247

Examples 247-257 are exemplary compounds of the invention having Formula(1) comprising 3-alkyl or 3-aryl substituted prolines, which may beprepared by repeating the procedures described in the above examples,using appropriate starting materials apparent to those skilled in theart.

Structure MS data 247

MS m/z 605.3 (M + 1) 248

MS m/z 667.4 (M + 1) 249

MS m/z 534.3 (M + 1) 250

MS m/z 596.4 (M + 1) 251

MS m/z 746.3 (M + 1) 252

MS m/z 698.3 (M + 1) 253

MS m/z 654.4 (M + 1) 254

MS m/z 716.4 (M + 1) 255

MS m/z 744.4 (M + 1) 256

MS m/z 682.4 (M + 1) 257

MS m/z 708.4 (M + 1)

Assays

The suitability of a channel activating protease inhibitor such as aprostasin inhibitor for the treatment of a disease mediated byinhibition of a channel activating protease, may be tested bydetermining the inhibitory effect of the channel activating proteaseinhibitor on: (1) the native, isolated, purified or recombinant channelactivating protease, using a suitable biochemical assay format, usingthe method described in Shipway et al.; Biochem. Biophys. Res. Commun.2004; 324(2):953-63); and/or (2) the ion channel/ion transport functionin suitable isolated cells or confluent epithelia, using the methodsdescribed in Bridges et al.; Am. J. Physiol. Lung Cell Mol. Physiol.2001; 281(1):L16-23; and Donaldson et al.; J. Biol. Chem. 2002;277(10):8338-45.

Biochemical Assays

Recombinant human prostasin and matriptase and guinea pig prostasin aregenerated according to methods described in Shipway et al., Biochem.Biophys. Res. Commun. 2004; 324(2):953-63). The recombinant enzymes areincubated in an electrolyte buffer containing the test compounds orvehicle in a suitable multiple well assay plate such as a 96 or 384 wellplate. At a defined time after the mixing of enzyme with compound orvehicle, a suitable fluorescent peptide substrate is added to the assaymixture. As substrate becomes cleaved by the active enzyme, fluorescence(measured, using a suitable fluorescence plate reader) increases and therate of turnover of substrate (i.e. enzyme activity) may be quantified,and thus the inhibitory effect of any test compound. The efficacy oftest compounds is expressed as the concentration that induces a 50%attenuation in the enzyme activity (K_(i)).

In general, compounds of the invention may have K_(i) values from 0.1 nMto 5 μM. In some examples, compounds of the invention may have K_(i)values from 0.1 nM to 500 nM; from 0.1 nM to 50 nM; from 0.1 nM to 5 nM;or from 0.1 nM to 0.5 nM. In particular examples, compounds of theinvention may have K_(i) values from 0.1 nM to 0.5 nM; from 0.5 nM to 5nM; from 5 nM to 50 nM; from 50 nM to 500 nM; or from 500 nM to 5 μM. Inyet other examples, compounds may have K_(i) values less than 0.1 nM ormore than 5 μM.

Epithelial Ion Transport

Human bronchial epithelial cells are cultured according to methodsdescribed in Danahay et al., Am. J. Physiol. Lung Cell Mol. Physiol.2002; 282(2):L226-36). When suitably differentiated (days 14-21 afterestablishing an apical-air interface) epithelial cells are treated witheither vehicle, aprotinin (200 μg/ml) or test compound for 90 minutes.Epithelia are then placed into, using Chambers as described in Danahayet al., Am. J. Physiol. Lung Cell Mol. Physiol. 2002; 282(2):L226-36)maintaining the concentration of vehicle, aprotinin or test compound onthe apical side of the epithelia. Short circuit current (ISC) is thenmeasured by voltage clamping the epithelia to zero millivolts. Theamiloride-sensitive ISC is then measured by the addition of amiloride(10 μM) to the apical surface of the epithelia. The potency of the testcompound is expressed as the concentration inducing a 50% inhibition ofthe total aprotinin-sensitive component of the amiloride-sensitive ISC.

In general, compounds of the invention may have IC₅₀ values from 1 nM to10 μM. In some examples, compounds of the invention may have IC₅₀ valuesfrom 1 nM to 1 μM; or more particularly from 1 nM to 100 nM. In yetother examples, compounds of the invention may have IC₅₀ values from 100nM to 1 μM, or from 1 μM to 10 μM. In yet other examples, compounds mayhave IC₅₀ values less than 1 nM or more than 10 μM.

Tracheal Potential Difference (In Vivo)

Guinea pigs are anaesthetized, using a short acting inhalationanaesthesia such as halothane and N₂O. While under short actinganaesthesia, an oral gavage needle is inserted into the trachea via theoropharangeal route. Once inside the trachea, a small volume (50-200 μl)of vehicle or test compound, in a suitable aqueous-based diluent, isinstilled into the airways. Animals then recover and become fullyambulatory. Alternatively, test compounds may be administered toanimals, using aerosol or dry powder dosing. At a defined time afterdosing, the animals are surgically anaesthetized, using a suitableanaesthesia such as ketamine and xylazine. The trachea is then exposedand a plastic agar bridge electrode is inserted into the tracheal lumen.A reference electrode is also inserted into the layers of muscle in theanimal's neck. The tracheal potential difference is then measured, usinga suitable high impedance voltmeter as described in Takahashi et al.,Toxicol Appl Pharmacol. 1995; 131(1):31-6. The potency of the testcompound is expressed as the dose inducing a 50% reduction in thesensitive-component of the tracheal potential difference.

It is understood that the examples and embodiments described herein arefor illustrative purposes only, and that various modifications orchanges in light thereof will be suggested to persons skilled in the artand are to be included within the spirit and purview of this applicationand scope of the appended claims. All publications, patents, and patentapplications cited herein are hereby incorporated by reference for allpurposes.

1. A compound of Formula (1):

or pharmaceutically acceptable salts, or stereoisomers thereof, wherein:J is a 5-12 membered monocyclic or fused carbocyclic ring, heteroaryl orheterocyclic ring containing N, O and/or S; R¹ is —(CR₂)_(l)—NR₂,—(CR₂)_(l)—NRC(═NR)—NR₂, —(CR₂)_(l)—C(═NR)—NR₂ or a 5-7 memberednitrogen-containing non-aromatic heterocyclic ring; W—R² is asubstituent at any position on ring A; W is —O(CR₂)_(k)—, —S(CR₂)_(k)—,—S(O)(CR₂)_(k)—, —SO₂(CR₂)_(k)— or —OC(O)(CR₂)_(k)—; R² is C₁₋₆ alkyl,C₂₋₆ alkenyl, C₂₋₆ alkynyl, R⁸, —CR¹⁰═CR¹⁰—R⁸, or

wherein ring E is an optionally substituted 5-7 membered monocyclic orfused carbocyclic or heterocyclic ring; or W—R² together form C₁₋₆alkyl, aryl or —OC(O)NR⁶R⁷; Y is SO₂R³ or SO₂NR⁶R⁷; R³ is C₁₋₆ alkyl,C₂₋₆ alkenyl, C₂₋₆ alkynyl, —(CR₂)_(l)—C₃₋₇ cycloalkyl or —(CR₂)_(l)—R⁸;R⁴ is —CR¹⁰═CR¹⁰—R⁸, —CR[(CR₂)_(l)—R⁸]₂, C₂₋₆ alkynyl, —O—(CR₂)_(l)—R⁹,NR⁶R⁷,

or an optionally substituted 5-7 membered carbocyclic ring, heterocyclicring, aryl or heteroaryl; or R⁴ together with Y form an optionallysubstituted 5-12 membered non-aromatic heterocyclic ring; R⁵ is halo,C₁₋₆ alkyl, C₁₋₆ alkoxy, OR⁹ or R⁹; R⁶ and R⁷ are independently H, C₁₋₆alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl or —(CR₂)_(l)—R⁸; or R⁶ and R⁷together with N may form an optionally substituted 5-7 memberedmonocyclic or fused heterocyclic ring; X, R⁸ and R⁹ are independently anoptionally substituted 5-7 membered carbocyclic ring, heterocyclic ring,aryl or heteroaryl; or R⁹ may be H or C₁₋₆ alkyl; R¹⁰ is H or C₁₋₆alkyl; each R is H, C₁₋₆ alkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl, wherein acarbon may optionally be substituted or replaced with NR, O or S; i is0-1; k, l and m are independently 0-6; n is 1-6; and p is 0-3.
 2. Thecompound of claim 1, wherein R¹ is —(CH₂)_(l)—NH₂,—(CH₂)_(l)—NHC(═NH)—NH₂ or —(CH₂)_(l)—C(═NH)—NH₂NH₂, wherein each l is0-1; or R¹ is piperidinyl.
 3. The compound of claim 1, wherein W is—O(CR₂)_(k)—, —S(CR₂)_(k)—, —S(O)(CR₂)_(k)—, —SO₂(CR₂)_(k)— or—OC(O)(CR₂)_(k)—; and k is
 1. 4. The compound of claim 1, wherein R² isan optionally substituted phenyl, thienyl, C₅₋₇ cycloalkyl, furanyl,piperidinyl, methylenecyclohexyl,


5. The compound of claim 1, wherein Y is SO₂R³; R³ is C₁₋₆ alkyl,—(CR₂)_(l)-cyclopropyl or —(CR₂)_(l)—R⁸ wherein R⁸ is an optionallysubstituted phenyl.
 6. The compound of claim 1, wherein R⁴ is —NH₂, oran optionally substituted phenyl, phenoxy, piperidinyl, C₅₋₇ cycloalkyl,cyclohexanol, imidazolyl, thienyl,

or R⁴ together with Y form an optionally substituted pyrrolidinyl,pyrrolidinonyl, tetrahydroisoquinolinyl or tetrahydronapthalenyl.
 7. Thecompound of claim 1, wherein -J-(R⁵)_(p) together is

Z is O or S; Z¹, Z², Z³ or Z⁴ are independently N, CH, or C whenattached to R⁵; Z⁵, Z⁶ or Z⁷ are independently N, O, S, CH, or C whenattached to R⁵; p is 0-1; and R⁵ is halo or C₁₋₆ alkyl.
 8. The compoundof claim 1, wherein R⁸ is an optionally substituted phenyl, C₅₋₇cycloalkyl, piperidinyl, cyclohexanol, imidazolyl, thienyl, furanyl,


9. The compound of claim 1, wherein i is 1; and X is an optionallysubstituted cyclohexyl, phenyl or piperidinyl.
 10. The compound of claim1, wherein J is benzothiazolyl, benzoxazolyl, thiazolyl, or oxadiazolyl.11. The compound of claim 1, wherein said compound has Formula (2A) or(2B):

wherein Z is O or S; R¹ is NH₂, —NHC(═NH)—NH₂ or —C(═NH)—NH₂; W is—O(CH₂)_(k)— or —S(O)(CH₂)_(k)—; R² is an optionally substituted phenyl,or W—R² together form C₁₋₆ alkyl or an optionally substituted phenyl; Yis SO₂R³; R³ is C₁₋₆ alkyl, —(CH₂)_(l)-cyclopropyl or —(CH₂)_(l)—R⁸wherein R⁸ is an optionally substituted phenyl; R⁴ is an optionallysubstituted, phenyl, piperidinyl, C₅₋₇ cycloalkyl, cyclohexanol,imidazolyl, thienyl,

i and p are 0; k is 1; l is 0-1; m and n are independently 1-4; and Rand R⁵ are as defined in claim
 1. 12. The compound of claim 11, whereinR⁴ is piperidinyl.
 13. The compound of claim 1, wherein said compoundhas Formula (3A) or (3B):

wherein R⁶ and R⁷ are independently H, C₁₋₆ alkyl or —(CR₂)_(l)—R⁸; orR⁶ and R⁷ together with N form an optionally substituted pyrrolidinyl,piperidinyl, morpholino, piperazinyl or diazepanyl; R⁸ is an optionallysubstituted phenyl, furanyl, tetrahydrofuranyl, piperidinyl or thienyl;and i and p are 0; and R, R¹, R⁵, X, Y, Z, i, l, m, n and p are asdefined in claim
 1. 14. A pharmaceutical composition comprising atherapeutically effective amount of a compound of claim 1 and apharmaceutically acceptable excipient.
 15. The compound of claim 1,wherein said compound is selected from the group